Weekly news updates on www.cli-online.com | October 2009 | Volume 33 | Issue 5 Protein profiling of arterial thrombosis in acute MI Pg.12 Turbidimetric NGAL test for automated systems Pg.34 Bottle top dispensor with safety discharge system Pg.37 Also in this issue : Reagent for measuring hyaluronic acid in clinical chemistry analysers Pg.41 О± thalassaemia screening Pg 8 NMR in clinical microbiology Pg 16 Markers of bone remodelling Pg 26 Setting the standard in allergy and autoimmunity diagnostics For accuracy and efficiency, run your allergy and autoimmunity blood samples on the worldвЂ™s leading test platform: ImmunoCAPВ®. Results are precise, correct and clinically reliable. Moreover, physicians trust them. Allergy and asthma ImmunoCAPВ® is an allergen-specific IgE antibody test that measures the exact levels of antibodies in the blood. By many considered the perfect tool for the diagnosis, prognosis and follow-up of patients with indications of allergy and asthma. Quantitative ImmunoCAPВ® tests can help the physician confirm or rule out allergy with certainty. Autoimmunity EliAв„ў is the premium autoimmunity diagnostic test, offering accuracy, standardization and guidance in a difficult diagnostic area. Clinically relevant and exact measurements of specific markers for several different autoimmune conditions, such as rheumatoid arthritis and celiac disease, are available. ImmunoCAPВ® brings accuracy, efficiency and economic value to laboratories. www.phadia.com www.cli-online.com & search 24594 We reflect passion and results in all we do. So do our award winners. Diane Davis, MT(ASCP)SH 1 3 1 Giacinto Gervasi, MS, MT(ASCP) 2 Cesare Manotti, MD 3 All ChildrenвЂ™s Hospital, St. Petersburg, FL. 2Syosset Hospital, Syosset, NY. Italian Federation of Centers for the Surveillance of Anticoagulant Therapies, Milan, Italy Congratulations to Diane Davis, Giacinto Gervasi, and Dr. Cesare Manotti, the 2009 global winners of ILвЂ™s вЂ�Passion & Results AwardвЂ™. In celebration of our 50th anniversary, IL is proud to honor three inspiring individuals who delivered remarkable results to improve patient care through their passion for diagnostic medicine. Like our winners, we, at IL, are driven by passion and results. Ours is an unmatched commitment to elevate patient care through our passion for developing the most innovative instruments, reagents and software, which produce the highest quality results. We salute the achievements of our three honorees and those of healthcare providers around the world, dedicated to delivering the best possible patient care. To learn about the accomplishments of our award winners, visit www.ilus.com/50forward. www.cli-online.com & search 24834 1 9 5 9 вЂ“ 2 0 0 9 Imagine ... innovating the science of histopathology Dedicated to Histopathology Sakura Finetek, again, improves the laboratory. By offering products to automate manual procedures and smoothen the workflow, the histotechnologists can easily complete the other activities required and eliminate potential risks. As the innovative company in histopathology, Sakura Finetek is continuously looking for possibilities to improve the laboratory... and succeeds in offering solutions for the problems found in the histopathology laboratory. Hall 3 D64 Sakura Finetek offers you the only concept to achieve: вЂў An efficient, manageable, continuous workflow вЂў Same day results by real reduction of turn-around time вЂў Higher productivity resulting in a higher morale вЂў Consistent high quality; sample by sample вЂў Improved health and safety standards in your laboratory First we understand. Then we innovate. Sakura Finetek Europe B.V. The Netherlands Phone: +31 71 589 83 00 Fax: +31 71 589 84 88 [email protected] www.sakura.eu www.cli-online.com & search 24727 08_009_SFE_5500_add-xSeriesA&P_D1 1 04-11-2008 16:15:29 Contents Biolyzer 200 В® FRONT COVER Weekly news updates on www.cli-online.com | October 2009 | Volume 33 | Issue 5 Protein profiling of arterial thrombosis in acute MI Pg.12 Turbidimetric NGAL test for automated systems Pg.34 The fine details of the molecular and cellular mechanisms underlying plaque rupture and thrombus formation in acute myocardial infarction are not yet fully elucidated. Profiling of cellular and soluble proteins from the arterial thrombus site may identify local effectors that amplify the vascular occlusion process illustrated on the front cover. The turn-key solution for the routine clinical laboratory Bottle top dispensor with safety discharge system Pg.37 Also in this issue : Reagent for measuring hyaluronic acid in clinical chemistry analysers Pg.41 О± thalassaemia screening Pg 8 NMR in clinical microbiology Pg 16 Markers of bone remodelling Pg 26 FEATURES [8-10] ble Now availa Rapid and accurate a-thalassaemia screening using quantitative real-time PCR [12-14] Protein profiling of arterial thrombosis in acute myocardial infarction [16-18] Applications of magnetic resonance spectroscopy in infectious diseases diagnosis [20-23] Rapid screening for antibiotic-resistant bacteria [24] ase study: blood film review C procedures in a general hospital lab [26-30] FOCUS ON FINLAND [26-28] N ovel biochemical markers of bone remodelling in the manage ment of osteoporosis [29] T he Finnish diagnostics industry: company profiles [30] Product news [34-38] Medica preview пѓњ Space efficient fully automated CC analyzer пѓњ B road field of application пѓњ Highly reliable and efficient system пѓњ Excellent customer support REGULARS [7] EditorвЂ™s letter [31-32] News in brief Medica Meet us at 3, in hall ber 3E67 m u n th o o b [40-41] Product news Analyticon Biotechnologies AG www.analyticon-diagnostics.com agile - affordable - accurate [42] Book reviews www.cli-online.com & search 24839 Salmonella E. coli 0157 Shigella Campylobacter Introducing the new Gold Standard test for screening of faecal pathogens. All within 24 hours. For the first time, the EntericBio system combines conventional microbiology and molecular techniques to produce accurate, efficient results. EntericBio is both unique and state of the art. For more information and a full demonstration video visit: EntericBio.com Serosep Limited Unit F5, Eastway Business Park, Ballysimon Road, Limerick, Ireland. Tel: +353 61 440 207 Email: [email protected] www.Serosep.com www.cli-online.com & search 24763 4 pathogens detected on 1 strip: > > > > Salmonella E. coli 0157 Shigella Campylobacter EditorвЂ™s letter 7 вЂ“ Issue NВ°5 вЂ“ October 2009 A society for all ages? The first of October marked the 10th anniversary of the UNвЂ™s International Day of Older Persons, and UN Secretary-General Ban KimoonвЂ™s message emphasised the need for persons over 60 (currently twenty percent of Europeans fall into this category) to be вЂ�both agents and beneficiaries of developmentвЂ™. On the same day a robust study led by danish professor Kaare Christensen was published in The Lancet, which concluded that if the increase in life expectancy in developed countries over the last two centuries is sustained in this century, over half of the babies born since the year 2000 will reach the age of a hundred. The ageing of the Western population has a serious impact on the old-age dependency ratio (the number of retired people divided by the number of working age people). This ratio is expected to double in forty years, greatly adding to the burden of the working population if action is not taken. A frequently suggested but facile solution to this complex problem is to raise the legal age of retirement from work. However, given that the average age of European menвЂ™s exit from the workforce ranges from 58.6 years in Belgium to 64.2 in the UK (in women it ranges from 58.2 in Austria to 62.9 in Romania) and that the official retirement age is 65 in most European countries, it must first be ensured by healthcare providers, employers and indeed society as a whole that the older workforce is healthy enough to continue in employment, at least until the legal retirement age. The most important condition for older employees to work longer (and be agents of development) is good health. Because the capacity for bearing weights decreases as workersвЂ™ age increases, musculoskeletal disorders due to carrying hefty loads or using heavy tools are the most frequent causes of work-related disability in older workers. Older persons are also more prone to work-related upper limb problems caused by awkward posture, repetitive movements and too few rest periods. In addition, work-related stress, predominantly caused by the demands being made exceeding the workerвЂ™s ability to cope without excessive working hours, is more frequent in older workers. Perceptive employers can do much to alleviate these problems, but commuting older workers face an additional hazard: they are less able to balance when standing in crowded commuter trains and buses. TodayвЂ™s older workers gave up their seats for elderly users of public transport when they themselves were young; if todayвЂ™s young people cannot be persuaded to emulate them, the provision of special seats for older commuters is necessary. People in the West are not only living longer, but have fewer disabilities and functional limitations than in the past, largely because they are the beneficiaries of development via healthcare systems offering effective diagnosis and treatment. However, if health services are not to be overwhelmed by an ageing population, it is vital that both workers and those who have retired from the workforce know how to safeguard their own health. An active life-style with regular exercise, a healthy diet, no smoking, limited use of alcohol and maintaining an appropriate blood pressure will hopefully allow many of todayвЂ™s infants to celebrate a happy and healthy 100th birthday. 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Via G. Mascherpa 14 20048 Carate Brianza (MB) Italia Fon +49 (0) 2534 8008-0 Fax +49 (0) 2534 8008-90 [email protected] Fon: 856 467 0018 Fax: 856 467 0188 US toll-free: 888 808 0067 [email protected] Fon +81 (0) 29 834 7788 Fax +81 (0) 29 834 7772 [email protected] Fon +33 (0) 1 69 04 87 12 Fax +33 (0) 1 69 04 90 38 [email protected] Fon +44 (0) 1227 823744 Fax +44 (0) 1227 824038 [email protected] Fon +39 0362 909 143 Fax +39 0362 909 157 [email protected] www.cli-online.com & search 24824 Disea se foc us вЂ“ Issue NВ°5 вЂ“ October 2009 8 Haemoglobinopathies Rapid and accurate a-thalassaemia-screening using quantitative real-time PCR technology Alpha-thalassaemia, a common genetic disorder leading in its most moderate form to a hypochromic microcytic anaemia, is mainly the result of deletions on chromosome 16. Current testing for a-thalassaemia is based on an algorithm of exclusion-testing, which necessitates a wide range of dianostic procedures. Such procedures are labourintensive, time-consuming and not 100% specific. As a result, there is a need for a general, rapid and efficient screening method. This article discusses the use of real-time quantitative PCR detection of deletions as one possible solution. by Dr S. Brunner-Agten and Dr A. R. Huber Alpha-thalassaemia a-thalassaemia is a common genetic disorder, which leads in its most moderate form to an asymptotic anaemia with persistent hypochromia microcytosis. The clinical outcome of more severe cases leads to very severe transfusion-dependent anaemia or hydrops fetalis. The condition is mainly the result of deletions on chromosome 16, which contains, at its telomeric region, two highly homologous and closely linked genes (al- and a2-gene) encoding the a globin chains. During meiosis, misalignment of chromosome homologues followed by reciprocal recombination at three highly homologous segments (named X, Y, and Z and separated by non-homologous segments [Figure 1]), results in various deletion-duplication events. Figure 2. Selected genotypes and corresponding ratio patterns. The outcomes of the genetic disorder are diverse and the severity is correlated with the number of affected a globin loci and the exact nature of the gene deletion [Figure 2]. is the result of the deletion or dysfunction of three of the four a-globin alleles (--/-a). It is characterised by microcytic hypochromic haemolytic anaemia, hepatosplenomegaly, mild jaundice and sometimes thalassaemialike bone changes. The phenotypes of a-thalassaemia represent two clinically significant forms, which are Hb Bart hydrops fetalis (Hb Bart) syndrome and haemoglobin H (HbH) disease. In HB Bart, all four a-globin alleles are deleted or inactivate (--/--) and death in the prenatal or neonatal period is inevitable. HbH, however, The phenotypes of a+-thalassaemia and a0-thalassaemia, where just one or two a-globin-genes are affected, are more common. a+-thalassaemia results from deletion or dysfunction of one a-globin allele (a a/-a), e.g. by reciprocal recombination between the Z region, 3.7 kb apart, or between the X Figure 1. Diagram of the a-globin gene cluster. Black boxes: highly homologous regions, separated by non-homologous segments; white boxes: exons encoding the a-globin chains. region, 4.2 kb apart, giving rise to the -a3.7kb and -a4.2kb deletion, respectively [Figures 1 and 2]. Carriers of a+-thalassaemia, also known as a-thalassaemia silent carrier, may have a silent haematological phenotype or present a moderate thalassaemia-like haematological picture. aВє-thalassaemia however, may be caused by extended deletions varying from 100 kb to more than 250 kb resulting in deletion or dysfunction of two a-globin alleles (homozygotes (-a/-a) or heterozygotes (a a/--), e.g. -a SEA, -a TAI, -a FIL, -a MED, -(a)20.5kb. Carriers of aВє-thalassaemia, also known as a-thalassaemia trait, show microcytosis (low MCV), hypochromia (low MCH) and normal percentages of HbО‘2 and HbF [1]. It is estimated that there are at least 200 million people worldwide affected by thalassaemia. In Switzerland, after iron deficiency, thalassaemia is the most prevalent cause of hypochromic anaemia [2, 3]. To offer genetic counselling for couples who wish to start a family, or to avoid unnecessary iron substitution, it is important to also identify the heterozygote carriers of a-thalassaemia who have mild or even no symptoms. 9 Available methods for alpha-thalassaemia screening Current testing for a-thalassaemia is based on an algorithm of exclusion-testing (i.e. to exclude iron deficiency and ОІ-thalassaemia and other haemoglobinopathies), which requires a wide range of procedures such as hematological testing of red blood cell indices, peripheral blood smears, supravital staining to detect RBC inclusion bodies, qualitative and quantitative haemoglobin analysis, bone marrow examination, and the in vitro synthesis of radioactively-labelled globin chains in affected individuals. However, the final proof of the presence of an a-thalassaemia is only obtained using biomolecular diagnostics [2]. This includes polymerase chain reaction (gap-PCR) amplification of the normal or aberrant a-globin gene [4,5], ELISA for the detection of aвЂ“globin chains in circulation [6] and hybridisation assays with a-strips. Current technologies are, however, labourintensive and time-consuming, and may still not provide an accurate analysis of all variants of the diseases. There is a great need for a general, rapid and efficient screening method, which is completely standardised and suitable for the routine laboratory. Quantitative real-time PCR as an a-thalassaemia screening method Real-time Quantitative PCR technique (RTPCR) has been applied in different investigations including pathogen detection, allelic discrimination, gene expression and gene regulation [7-9], as well as for the detection of duplications and deletions, e.g. in Duchenne and Becker muscular dystrophies, cystic fibrosis and neuroblastomas [10-12]. However, while RT-PCR has also been applied for the detection of a-thalassaemia [13], current methods only allow for detection of several restricted mutations such as the southeast Asian type deletion, or a group of three different deletions (-a 3.7kb, -a SEA and -a MED). We are now evaluating a new screening assay (patent pending) for the detection of a-thalassaemia-causing deletions using multiple primer sets, which enables classification of the genotype of an individual by performing only one (or maximally two) single RT-PCR run. In order to carry out the assay, genomic DNA from human blood is extracted using a manual or automated DNA purification method. Photometric quantification of genomic DNA is performed on the NanoDrop liquid handling device (Thermo Fisher Scientific, Inc.) and вЂ“ Issue NВ°5 вЂ“ October 2009 only samples within a defined range of DNA purity (260nm: 280nm ratio) are selected for the experiments. A Light Cycler System is used for RT-PCR. The specificity of the obtained amplicons is analysed through melting curves, gel electrophoresis and/or sequencing. Further quantification in reference to endogenous controls (reference genes) allows identification of the relative quantity of the amplified gene. Through analysis of the obtained amplification pattern we are able to define the genotype of the individual. In the case of an aberrant genotype (positive screening result), subsequent analysis, e.g. sequencing, allows further characterisation of the exact nature and location of the mutation (if this is relevant information for the clinic). In the case of negative screening results, no additional work-up in the alpha-gene-cluster is necessary. Conclusion With this quantitative RT-PCR assay we have developed a new, completely standardised method for routine laboratory alpha-thalassaemia-screening, which enables the genotype of each patient to be classified by performing one single RT-PCR run. The implementation of this new method in a diagnostic laboratory requires the assessment www.cli-online.com & search 24609 вЂ“ Issue NВ°5 вЂ“ October 2009 of a new algorithm for testing. The two first steps normally carried out, namely the determination of the haemogram and the iron metabolism parameters, would be retained. However, after iron deficiency and b-thalassaemia and haemoglobinopathies are excluded, the new a-thalassaemias screening method can be used instead of the common molecular biological tests used to date. The quantification of the a-globin gene will allow the determination of the real prevalence of a-thalassaemia, with the detection of all carriers who may otherwise be subject to mis- or nondiagnosis. 10 Haemoglobinopathies This helps to provide genetic advice and (prenatal) diagnostics. Beyond this, the method can help to minimise unnecessary, potentially toxic and expensive iron substitution in patients who have a-thalassaemia, rather than iron deficiency anaemia. References 1. Herklotz R, Risch L and Huber AR. Hemoglobinopathies--clinical symptoms and diagnosis of thalassemia and abnormal hemoglobins. Ther Umsch 2006; 63(1): 35-46. 2. Huber AR et al. Anomales HГ¤moglobin: Erscheinungsbilder und AbklГ¤rung. Swiss Medical Forum, 2004. 3. Huber AR et al. ThalassГ¤mie-Syndome: Klinik und Diagnose. Swiss Medical Forum, 2004. 4. Chang JG et al. Rapid diagnosis of alpha-thalassemia-1 of southeast Asia type and hydrops fetalis by polymerase chain reaction. Blood 1991; 78(3): 853-4. 5. K o TM et al. Carrier detection and prenatal diagnosis of alpha-thalassemia of Southeast Asian deletion by polymerase chain reaction. Hum Genet 1992; 88(3): 245-8. 6. Ausavarungnirun R. et al. Detection of zeta-globin chains in the cord blood by ELISA (enzymelinked immunosorbent assay): rapid screening for alpha-thalassemia 1 (Southeast Asian type). Am J Hematol 1998; 57(4): 283-6. 7. B owie LJ et al. Detection of alpha-thalassemias by multiplex polymerase chain reaction. Clin Chem 1994; 40(12):2260-6. 8. D as H et al. Quantitation of Fas and Fas ligand gene expression in human ovarian, cervical and endometrial carcinomas using real-time RT_PCR. Br J Caner 2000; 82(10): 1682-8. 9. F ujii K et al. Mutation detection by TaqMan-allele specific amplification: application to molecular diagnosis of glycogen storage disease type Ia and medium-chain acyl-CoA dehydrogenase deficiency. Hum Mutat 2000; 15(2): 189-96. 10. J ancourt F et al. Rapid identification of female carriers of DMD/ BMD by quantitative real-time PCR. Hum Mutat 2004;. 23: 385вЂ“391. 11. S chneider M et al. Detection of exon deletions within an entire gene (CFTR) by relative quantification on the LightCycler. Clin Chem 2006; 52(11): 2005-12. 12. DePeter K et al. Quantification of MYCN, DDX1, and NAG gene copy number in neuroblastoma using a real-time quantitative PCR assay. Mod Pathol 2002; 15(2): 159-66. 13. Armour JA et al. The detection of large deletions or duplications in genomic DNA. Hum Mutat 2002; 20(5): 325-37. The authors Dr Saskia Brunner-Agten and Prof. Andreas R. Huber Cantonal Hospital Aarau (KSA) Switzerland Address for correspondence: Prof. Dr. med. Andreas R. Huber Zentrum fГјr Labormedizin Kantonsspital Aarau AG Tellstrasse CH-5001 Aarau Switzerland Tel :+41 62 838 53 01 E-mail: [email protected] Comments on this article? Feel free to post them at www.cli-online.com/comment/alpha_thalassaemia The coinheritance of beta- and alpha- thalassaemia: a review of one patient and her family The diagnosis and management of alpha-thalassaemia may be complicated by the variability of the phenotype, which is due to the interaction of coinherited alpha-thalassaemia and the variable severity of beta-thalassaemia mutations. A well-documented case of complex beta- and alpha-thalassaemia coinheritance is described in this paper. Laboratory and clinical data for the patient and her family were reviewed. The patient was an asymptomatic girl, one of identical twins. She presented at one month of age for follow-up of an abnormal newborn-screening result (haemoglobin F only), which initially suggested homozygosity for beta-thalassaemia. Extensive studies on the patient and family revealed that she had coinherited alpha-thalassaemia traits and homozygous beta-thalassaemia. This case demonstrates the interaction of coinherited alpha- and betathalassaemia with the resultant amelioration of the clinical phenotype. It also highlights the importance of family studies and close follow-up in diagnosing complex haemoglobinopathies. Mast KJ, Hammond S, Qualman SJ, Kahwash SB. Lab Hematol. 2009;15(3):30-3. www.cli-online.com & search 24830 В© 2008 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries. Champion of ergonomics, efficiency and endurance. Introducing the new Thermo Scientific Finnpipette F2, a pipette that has all the right qualities вЂ“ it has a light pipetting action, itвЂ™s easy to use, and itвЂ™s fully autoclavable. Add to this great accuracy В® and precision, and you know the Finnpipette F2 is a winner. вЂў New Advanced Volume Gearing improves accuracy and precision вЂў New large digit display вЂў Very light and smooth pipetting action вЂў Fully autoclavable вЂў 5 year warranty To order the new Thermo Scientific Finnpipette F2 or for more information contact your local representative or visit www.thermo.com/finnpipette. Visit us at Medica 2009 Dusseldorf/Germany вЂ“ November 18-21 Hall 01 / Both E11 Moving science forward www.cli-online.com & search 24852 The ideal system for good laboratory pipetting. The Finnpipette F2 combines with Thermo Scientific FinntipВ® Flex pipette tips to form a complete, optimized pipetting system. M ol ec ul a r d i a g nost i c s вЂ“ Issue NВ°5 вЂ“ October 2009 12 Cardiovascular disease Protein profiling of arterial thrombosis in acute myocardial infarction Atherosclerotic plaque rupture with subsequent mural thrombus formation is considered the main event compromising epicardial flow in acute myocardial infarction (AMI). In the past, only a few selected biomarkers have been targeted to characterise the complex and poorly understood mechanisms underlying acute coronary occlusion. Recent proteomic techniques now enable a comprehensive analysis of protein alterations during atherothrombosis. As the main pathophysiological determinants, proteins condition the propensity and progression of atherothrombosis and cardiovascular disease. In particular, profiling of cellular and soluble proteins from the arterial thrombus site may identify local effectors that amplify the vascular occlusion process in AMI. by Dr M. Kubicek, Dr K. Distelmaier and Dr I. M. Lang Thrombus formation in acute myocardial infarction Thrombotic occlusion of an epicardial coronary artery upon atherosclerotic plaque rupture is considered the ultimate and key step in acute myocardial infarction (AMI) [1]. The propensity of vulnerable plaque rupture and subsequent thrombus formation depends on a complex cascade of events involving endothelium, inflammatory cells, cytokines, endothelin, complement system, apoptosis, T-lymphocytes, metalloproteinases, phospholipase A2, cholesterol, and platelets [2]. However, the molecular and cellular mechanisms underlying plaque rupture and thrombus formation in AMI are not fully elucidated. Timely recanalisation of the occluded artery by percutaneous coronary intervention (PCI) [Figure 1] is critical to limit cardiac mortality and morbidity [3]. Implantation of drug eluting stents during PCI is carried out to reduce re-stenosis and re-occlusion of the culprit artery, which harbours the potential for late thrombosis, thus compromising long-term prognosis [4]. Impaired re-endothelialisation and local inflammatory reactions may contribute to stent-thrombosis. In the past, the basic knowledge of local processes underlying plaque rupture and thrombus formation has mainly been derived from post-mortem material obtained hours after the event, or from animal model systems [5]. Figure 1. Schematic diagram of sample collection (A) and processing (B) during PCI of acute myocardial infarction patients. In C-E the culprit lesion is made visible (arrows indicate beginning of the thrombotic occlusion site and the distal position of the aspiration catheter). With the advent of modern aspiration catheters, fresh plaque, particulate thrombus material and also whole blood can be harvested from the site of coronary occlusion during primary PCI and compared to material aspirated at systemic sites of the same patients [Figure 1]. Several studies have investigated local concentrations of selected molecules at the culprit lesion site [6-8]. Proteomic approaches to analyse thrombosis in AMI Changes in the abundance or structure of plasma or cellular proteins at the thrombus site can directly alter clot morphology and function and induce downstream events relative to patient outcome. Recent technical achievements in liquid chromatography/mass spectrometry (LC-MS/MS) facilitate the global profiling of proteins and selected markers involved in arterial thrombosis. In the following article, we provide an overview of current proteomic studies on relevant samples. Proteome profiling of blood plasma Direct proteomic analysis of whole plasma from healthy individuals, unstable angina or AMI patients has revealed differences in relatively highly abundant plasma proteins such as alpha-1 antitrypsin, apolipoprotein A-1 and fibrinogen Оі-chain [9], as well as AMI specific proteolysis patterns (i.e. complement C3f and fibrinopeptide A) [10]. Affinity depletion of high abundant proteins enables access to medium to low abundant plasma proteins, but may also co-deplete disease relevant proteins from the sample [11]. A comprehensive analysis of immunodepleted plasma with differential LC-MS/MS identified that 95 out of 731 proteins differed between individuals with or without angiographic coronary artery disease (CAD) [12]. One approach to identifying less abundant thrombosis-related proteins is to assess plasma derived directly from the site of plaque rupture rather than from peripheral blood. We recently reported two-dimensional gel electrophoresis (2d-GE) of whole plasma generated from the culprit or the systemic site of AMI patients undergoing PCI [13]. We showed a local accumulation of complement effector proteins that are involved in neutrophil recruitment to the culprit lesion site. Multiple immunodepletion 13 of the same samples followed by both 2d-GE and LC-MS/MS identified local accumulation of additional complement effectors and regulators [13] but also of a novel factor (personal unpublished data). This low abundant protein was shown to be specifically expressed and processed at the thrombus site in vitro and in vivo, and was proteomically not detected in peripheral blood. Proteome profiling of particulate thrombus material Particulate thrombus is mainly constituted by cross-linked fibrin, but also contains a number of proteins and cells that may alter clot structure and function. Howes et al analysed clot-associated proteins by 2d-GE and LC-MS/MS [14]. Our own group has identified 695 different protein species in particulate thrombus material harvested during PCI from AMI patients (personal unpublished data). Screening of a self-designed proteomic database (www.meduniwien.ac.at/proteomics/database) [15] for cell type-specific and functional signatures revealed neutrophil-specific effector functions in particulate thrombus material. Interestingly, the proteome of late STENT-induced clots differed markedly from that of native thrombi when compared by spectral counting of shotgun LC-MS/ MS data (personal unpublished data). We hope that these proteomic data will help to elucidate the specific mechanisms of STENTinduced late thrombosis and help to prevent these life-threatening complications in the future. Proteome profiling of circulating cells Circulating blood cells like platelets or leukocytes can interact with the vasculature and the thrombus at the culprit lesion site and, upon activation, express specific adhesion molecules, secrete chemokines and cytokines, liberate the contents of alpha granules, generate reactive oxygen species (ROS) and release microparticles (MP). Changes in protein expression and phosphorylation of platelets have been studied upon in vitro activation with thrombin [16], thrombin receptor activating peptide (TRAP) [17], or collagen-related peptide [18]. More than 300 different proteins were found to be released by platelets following thrombin activation by a combination of 2d-GE-based and LC-MS/MS [19]. Several of the identified proteins were not previously assigned to platelets, including calumenin and secretogranin 3, and were found expressed in atherosclerotic but not healthy arteries. Thrombogenic proteins have been identified, including platelet factor 4 (PF-4), associated with MP released from ADP activated platelets [20]. Barderas et al have analysed circulating peripheral blood monocytes isolated from patients with acute coronary syndrome (ACS) or stable CAD by 2d-GE and LC-MS/MS [21]. The authors вЂ“ Issue NВ°5 вЂ“ October 2009 identified several intracellular proteins that were altered in the acute phase of ACS, changes which were reversed to the level of stable patients after a six month period post-admission. To date no proteome profile of monocyte secretomes or monocytes from the culprit lesion site have been reported. We have recently demonstrated that polymorphonuclear neutrophils (PMNs) accumulate at Your Power for Health Power of Caring MiniCollectВ® Capillary Blood Collection for Little Patients Gentle & comfortable collection for children and neonates Unique вЂњCross-Cuts CapвЂќ means no need to decap tubes Closed system for maximum safety Flexible system for use with funnel or capillary Greiner Bio-One GmbH | Bad Haller StraГџe 32 | A-4550 KremsmГјnster Phone: (+43) 75 83 67 91-0 | Fax: (+43) 75 83 63 18 | E-mail: [email protected] www.gbo.com/preanalytics www.cli-online.com & search 24516 вЂ“ Issue NВ°5 вЂ“ October 2009 14 the site of plaque rupture in AMI, and that the number of accumulated PMNs correlates with ST-segment resolution [22] and enzymatic infarct size [13]. Secreted proteins of activated PMNs have been mapped by 2d-GE- and 2D-LC-based approaches identifying thrombomodulatory factors such as cathepsin G or calreticulin [23-24]. We have assessed the proteome of PMNs harvested from the culprit lesion site in AMI by 2d-GE and LC-MS/MS (personal unpublished data). In addition, we have employed metabolic in vitro labelling with 35S methionine/cysteine prior to autoradiography of 2d-gels generated from PMN cytosols and supernatants [Figure 2]. This approach allows the identification of low abundant cytokines, chemokines and growth factors from cell supernatants [25] while contaminating proteins from plasma, platelets or erythrocytes are not labelled [26]. We have identified reproducible proteomic alterations in PMNs incubated with plasma generated from the culprit lesion site versus the systemic site [personal unpublished data; see Figure 3]. These low abundant proteins can in turn be assayed in clinical specimens by high sensitive methods. Conclusions Atherothrombosis is one of the major causes of morbidity and mortality. Identification of the underlying molecular mechanisms is needed to improve cardiovascular risk and prognosis. Modern proteome profiling approaches from material harvested at the thrombus site in AMI have a high potential to identify important effector proteins implicated in atherothrombosis. In parallel, in vitro stimulation of relevant primary cell types allows the capture of low abundant proteomic changes. Extended application of proteomic profiling to different determinants of plaque rupture and thrombosis will enable a comprehensive assessment of the complex molecular processes relevant to atherothrombosis and acute vascular syndromes. References 1. Naghavi M et al. Circulation 2003;108(14). 2. Libby P. J Intern Med 2008;263:517-27. 3. D e Luca G et al. J Am Coll Cardiol 2003;42:991-7. Cardiovascular disease Figure 2. Schematic diagram of 2d-GE prepared from in vitro labelled primary circulating blood cells. 4. Inoue T et al. Circ J 2009;73:615-21. 5. Cullen P et al. Arterioscler Thromb Vasc Biol 2003;23:535-42. 6. Maeda N et al. Fundam Clin Pharmacol 2009;23:351-7. 7. Maier W et al. Circulation 2005;111(11). 8. Suzuki M et al. Angiology 2006;57:459-63. 9. Mateos-Caceres PJ et al. J Am Coll Cardiol 2004;44(8):1578-83. 10. Marshall J et al. J Proteome Res 2003;2(4) :361-72. 11. Stempfer R et al. Electrophoresis 2008;29(21) :4316-23. 12. D onahue MP et al. Am Heart J 2006; 152:478-85. 13. Distelmaier K et al. Thromb Haemost 2009;102:564-72. 14. Howes JM et al. Diab Vasc Dis Res 2008; 5:205-12. 15. Wimmer H et al. Electrophoresis 2009; 30:2076-89. 16. Maguire PB et al. Proteomics 2002;6:42-8. 17. Garcia A et al. Blood 2004 15;103:2088-95. 18. Garcia A et al. Proteomics 2006;6:5332-43. 19. Coppinger JA et al. Blood 2004;103:2096-104. 20. G arcia BA et al. J Proteome Res 2005; Figure 3. Pseudocolour-overlay (fluorographs magenta; autoradiographs green) of cell supernatants generated from PMNs incubated with plasma obtained from the culprit lesion site (A, B) and the systemic site (C, D), respectively. Arrows indicate altered protein spots that were affected by a specific neutrophil inhibitor. 4:1516-21. 21. B arderas MG et al. Methods Mol Biol 20 07;357:319-28. 22. Adlbrecht C et al. Thromb Haemost 2007;97(4). 23. B oussac M et al. Electrophoresis 2000; 21(3):665-72. 24. L ominadze G et al. Mol Cell Proteomics 2005;4:1503-21. 25. G undacker N et al. Electrophoresis 2006; 27:2712-21. 26. H audek VJ et al. J Proteome Res 2009;8:3834-43. The authors Markus Kubicek*, Klaus DistelmaierВ§, and Irene M. LangВ§ *D epartment of Medical and Chemical Laboratory Diagnostics, Medical University of Vienna and В§ Department of Internal Medicine II, Division of Cardiology, Medical University of Vienna Corresponding author: Markus Kubicek Department of Medical and Chemical Laboratory Diagnostics Medical University of Vienna Waehringer Guertel 18-20 A-1090 Vienna, Austria Tel: +43 1 40400 6436 Fax: +43 1 40400 6437 E-mail: [email protected] Comments on this article? Feel free to post them at www.cli-online.com/comment/Protein_profiling If You Are Not Using Nova's Glucose StatStrip , You Are Misdosing Insulin Dependent Patients With Abnormal Hematocrits в„ў According to a recent published study1 by the Mayo Clinic, only Nova's StatStrip continued to give accurate glucose results as patient hematocrits varied from 20% - 65%. Hematocrit related errors of 20% to nearly 50% were reported on every glucose meter system tested except for Nova's StatStrip. Figure (b): Glucose 247 mg/dL StatStrip 15 (.83) 10 Accu-Chek PCx 10 (.56) Glucose Error Mean glucose difference (%) Glucose Error Mean glucose difference mg/dL (mmol/L) Figure (a): Glucose 54 mg/dL SureStep 5 (.28) 0 (0) -5 (-.28) -10 (-.56) -15 0 -10 -20 -30 Accu-Chek -40 (-.83) PCx SureStep -50 -20 (-1.11) 20 30 40 50 60 70 20 30 40 60 50 70 Hematocrit (%) Hematocrit (%) Figure (c): Glucose 483 mg/dL Figures: (a) Mean glucose difference (meter glucose minus reference glucose) and (b) and (c) mean glucose percent difference [(meter glucose minus reference glucose)/reference glucose x 100)] as a function of hematocrit at glucose concentrations of (a) 54 mg/dL, (b) 247 mg/dL, and (c) 483 mg/dL. Each point represents the mean В± standard deviation of the mean glucose difference or mean glucose percent difference (n=6). 10 Glucose Error Mean glucose difference (%) StatStrip 0 -10 -20 -30 StatStrip Accu-Chek -40 PCx SureStep -50 20 30 40 50 60 70 Hematocrit (%) Karon BS et al (Mayo Clinic) Evaluation of the Impact of Hematocrit and Other Interference on the Accuracy of Hospital Based Glucose Meters. Diabetes Technology and Therapeutics, April 2008 1. For further information, visit our website or contact us as indicated below. IN THE U.S., call toll-free 800-458-5813 or 781-894-0800 вЂў IN CANADA, call toll-free 800-263-5999 www.statstripglu.com www.cli-online.com & search 24762 La b t ec hnol og y вЂ“ Issue NВ°5 вЂ“ October 2009 16 NMR Applications of magnetic resonance spectroscopy in infectious diseases Nuclear magnetic resonance spectroscopy (NMR) is an analytical tool, widely used in physics, chemistry, material sciences, biochemistry, biomedical sciences and medicine. Applications in microbiology and for the diagnosis of infectious diseases are summarised in this article. by Dr Uwe Himmelreich Nuclear magnetic resonance spectroscopy (NMR) is a powerful tool for the identification of chemicals, mainly in solutions. By this means, magnetic properties of nuclei are used to gain information on the chemical structure of a molecule. The hydrogen atom (1H) is the most sensitive and widely used isotope in NMR spectroscopy. It can also be used to gain information on the distribution of molecules such as in magnetic resonance imaging (MRI). The application of NMR spectroscopy to biological samples such as cells, body fluids or biopsy samples provides information on a large range of metabolites. Due to the non-invasive nature of the technique, one is not restricted to tissue extracts but can also study aspects of cellular biochemistry in living systems. In vivo NMR spectroscopy is routinely used in the clinic, for example to diagnose and assess tumours, stroke, metabolic diseases and other pathologies [1]. Its attractiveness results from its capability of simultaneous recognition of up to hundreds of metabolites in living cells maintained under physiological conditions. NMR spectroscopy requires homogeneous magnets operating at high field strength (NMR spectrometer or MRI scanner). Figure 1 illustrates the relationship between magnetic field strength and the potential of NMR in clinical diagnosis. Depending on the nature of the sample (extracts, biopsies, small animals or humans), stronger or weaker magnets can be used. The value of such a non-invasive diagnostic test performed on a patient is higher than tests that require invasive methods such as the collection of tissue samples. The application of NMR spectroscopy in clinical microbiology and infectious diseases is still in its infancy. Conventional diagnosis of infectious diseases relies on microbiological tests based on microbial culture and/or a limited array of biochemical tests. Disadvantages include the timeconsuming nature of the tests, and occasional poor sensitivity and specificity. Newly emerged approaches include molecular methods, which are rapid and discriminatory but currently limited in scope (ie target specific pathogens), miniaturised high throughput biochemical techniques and immunological approaches. Technologies such as NMR, but also infrared and Raman spectroscopy, chromatography and mass spectrometry, generate complex data based on chemical composition of microorganisms, which can detect phenotypic differences in closely related species. The potential for adaptation of such platforms for diagnostic use has been facilitated by automation of sample delivery and analysis, and development of computer-based methods of data interpretation. Characterisation of metabolites isolated from microbial cultures NMR spectra from secondary metabolites and other compounds, such as proteins, lipids or carbohydrates isolated from respective groups of microorganisms, have been utilised since the 1970s for the classification and identification of bacteria, fungi and lichenised fungi [2]. Compounds isolated from the cell wall or capsule material were most frequently utilised, but analyses of relatively small compounds secreted from cell suspensions also reveal differences between groups of microorganisms. Those approaches were, however, time consuming and lacked the sensitivity to distinguish between closely related species. Identification of microorganisms based on NMR spectra of cell cultures Figure 1. Comparison of the performance of NMR spectroscopy on different samples By using cell suspensions instead of extracts, NMR spectroscopy can become a rapid Figure 2. Comparison of NMR spectra from pus of an abdominal abscess (D) and cell suspensions of bacteria isolated from the pus ((A) Bacteroides fragilis, (B) Streptococcus milleri, and (C) Escherichia coli). screening method. Although the spectral quality suffers substantially by using suspensions of microorganisms, it is still sufficient to distinguish between different pathogenic species. For example, linear discriminant analysis of NMR data obtained from cell suspensions of various taxa of bacteria is able to identify them rapidly with an accuracy greater than 95% [3]. More sophisticated analysis methods of NMR spectra are able to distinguished between two most common pathogenic yeast species [4]. The identification accuracy for five different Candida species was comparable to molecular identification methods, and in some cases superior to sophisticated traditional identification systems used in clinical microbiology laboratories. Even the identification to sub-species level was possible and robust for the pathogenic yeast Cryptococcus neoformans [5]. The ease of sample preparation, rapid automated identification, and the robust nature of combining NMR spectroscopy with 17 computerised data analysis methods are attractive for clinical and industrial applications. In practice, NMR-based identification may be of most value for bacterial and fungal species, which are relatively slow growing or difficult to identify by conventional methods. Antimicrobial susceptibility testing Due to an increasing number of microbial strains resistant to antibiotics, rapid drug susceptibility testing is critical for choosing the right therapy for infectious diseases. Since the exposure of a microorganism to an antimicrobial drug most likely also influences its metabolism, it can be expected that antimicrobial drug pressure would similarly exert measurable effects on the metabolism. This has been demonstrated by measuring Metabolic End Points (MEP) for several Candida strains exposed to various antifungal drugs (caspofungin, AMB and voriconazole) [6]. NMR can simultaneously monitor utilisation of substrates (glucose etc.) and metabolites (acetate, ethanol etc.) that were affected by the exposure to the drug. Clear-cut and reproducible MEPs, defined as a >50% change in metabolite or nutrient concentrations were correlated with minimal inhibitory concentrations determined by conventional tests. It was confirmed that metabolic changes were evident much earlier than drug-induced inhibition of growth, the standard endpoint currently used to determine susceptibility to drugs. This illustrates the potential of NMR spectroscopy to provide a more rapid reading and thus to allow selection of the most appropriate therapy earlier than by more labour intense conventional tests. NMR of biofluids and biopsy material Using isolated microorganisms for the diagnosis of infectious diseases is still very time consuming. Rapid identification of microorganisms without cultivation and isolation of the organisms is therefore a longstanding aim for microbiologists. The analysis of biofluids like urine, cerebrospinal fluid (CSF) or blood by NMR spectroscopy has a long database of NMR spectra from clintradition, in particular for toxico- ical specimens that would cover all logical studies (for example [7]). In possible infective microorganisms. contrast to applications in oncology or toxicology, where only two or a The feasibility of identification of few potential diagnoses are possi- infection-causing microorganisms ble (for example malignant versus directly from biofluids by using benign), applications of NMR spec- NMR spectroscopy has been demtroscopy for the identification of onstrated in proof-of-principle infection-causing microorganisms studies based on pus from abscesses from biofluids (CSF, blood, pus) is [8-10]. Those studies demonstrated more difficult due to the diversity that: (1) the differences in specof pathogens and the associated troscopic pattern depend on the pub LCI jib :Mise en page 1 05/10/09 17:45 Page1 difficulties of building up a large causative organisms rather than the вЂ“ Issue NВ°5 вЂ“ October 2009 site of sample collection, (2) main metabolites produced by bacteria in liquid culture are also dominant in pus samples, and (3) distinguishing between major groups of pathogens is feasible. Figure 2 shows the example of 1H NMR spectra from a human pus sample and bacterial cell suspensions of organisms isolated from the pus. Key bacterial metabolites such as organic acids (acetate, butyrate or succinate) and amino acid residues are also present in the NMR spectrum of pus. Gain de temps, contrГґle des coГ»ts, et gestion du flux de travail en toute sГ©rГ©nitГ© вЂ¦ Le 4 novembre 2009, vous dГ©couvrirez en exclusivitГ© HaemCell, une nouvelle vision de lвЂ™organisation du laboratoire dвЂ™hГ©matologie. vous Rendez- e Curie ari rre et M Hall Pie 5 Stand C4 www.horiba.com/medical www.cli-online.com & search 24476 вЂ“ Issue NВ°5 вЂ“ October 2009 18 NMR The feasibility of NMR spectroscopy for rapid, aetiological diagnosis of meningitis was demonstrated using as little as 10 ВµL CSF in animal models for meningitis [11]. Accurate distinction between sterile CSF, and meningitis caused by Streptococcus pneumoniae and Cryptococcus neoformans was possible. An agreement of 98% between the NMRbased classification and the microbiological diagnosis was achieved. The application of NMR for the diagnosis of infectious diseases based on biofluids is also promising due to the ease of automated, high-throughput data acquisition and analysis, and hence automated diagnosis of infections. The potential of any high throughput diagnostic platform, which will achieve maximum efficiency (cost/benefit) when operating continuously, is dependent on the number of samples of a given type and the versatility of the platform. Different types of sample (cultured cells, cell supernatants, biofluids, tissue biopsies) and different types of pathology (microbiology, clinical chemistry, tissue pathology) are readily analysed in an NMR laboratory, which makes such platforms potentially attractive for large hospitals. In vivo Magnetic Resonance Spectroscopy of brain infections Magnetic Resonance Imaging has increased the ability to localise cerebral lesions but is not able to provide a definite diagnosis. In patients with focal lesions, stereotactic biopsy or open surgical procedures are required to obtain tissue for histopathological and microbiological examination. In contrast, in vivo NMR spectroscopy may provide a non-invasive alternative. Figure 3 shows NMR images and spectra of an infective abscess compared to a brain tumour. An NMR spectrum obtained from the location of the lesion (indicated by the square box in the MRI) clearly distinguishes between the two pathologies. Identification of marker metabolites like acetate, succinate and lactate can be used as an indication to distinguish between infective abscesses and brain tumours non-invasively. Attempts were already made to classify infection-causing microorganisms based on in vivo NMR spectra [10, 12, 13]. However, aetiological identification of infective brain lesions using NMR spectra is not yet available due to the low number of reference cases. As clinical management of both diseases is different, in vivo NMR spectroscopy can optimise therapy and avoid unnecessary diagnostic procedures and surgery. Conclusion Microbiology laboratories in clinics, industry and agriculture must be able to identify a wide number of pathogens accurately and rapidly. Microbiological tests should be inexpensive, fully automated, not depend on specialist knowledge about the technology, be applicable to a Figure 3. In vivo MR images and NMR spectra from a brain abscess and tumour. Distinction between tumour (increased choline to creatine ratio) and abscess (presence of acetate and succinate) is possible based on marker metabolite signals. broad range of microorganisms and be as noninvasive as possible. Fully automated acquisition of NMR spectra is available in many industrial and pharmaceutical laboratories for screening of large quantities of samples. On the other hand, automated incubation of microbial cultures is a reality in many microbiological laboratories. Combining both approaches with automated data analysis and currently available robotic technology will result in a fully automated identification of microorganisms with high sample throughput, high accuracy and minimal manual handling and processing. However, the ultimate aim of future developments should be the non-invasive diagnosis of certain infections without the need of taking tissue samples. References 1. Danielsen ER & Ross BR. Magnetic Resonance Spectroscopy. Diagnosis of Neurological Diseases. Marcel Dekker Inc, New York, 1998. 2. Grivet JP, Delort AM, Portais JC. Biochimie 2003; 85: 823вЂ“840. 3. Bourne R, Himmelreich U, Sharma A et al. J. Clin. Microbiol 2001; 39: 2916-2923. 4. Himmelreich U, Somorjai RL, Dolenko B et al. Appl. Environ Microbiol 2003; 69: 4566-4674. 5. Sorrell TC, Wright LC, Malik R & Himmelreich U. FEMS Yeast Res 2006; 6: 558-566. 6. Coen M, Bodkin J, Power D et al. Antimicrob Agents Chemother 2006; 50: 4018-4026. 7. Lindon JC, Holmes, E & Nicholson JK. Prog NMR Spect 2004; 45: 109-143. 8. Garg M, Gupta RK, Husain M et al. Radiology 2004; 330: 519-527. 9. Himmelreich U, Dzendrowskyj T, Allen C et al. Radiology 2001; 220: 122-128. 10. Himmelreich U, Accursso R, Malik R et al. Radiology 2005; 236: 261-270. 11. Himmelreich U, Malik R, KГјhn T et al. PLoS One 2009; 4(4): e5328. 12. Agarwal M, Chawla S, Husain N et al. Neuroradiol 2004; 46: 211-215. 13. H immelreich U & Gupta RK. Application of magnetic resonance for the diagnosis of infective brain lesions. In: Webb GA(Ed.) Modern Magnetic Resonance 2006; 2: 991-999. The author Dr Uwe Himmelreich Biomedical NMR Unit MoSAIC Department of Medical Diagnostic Sciences Katholieke Universiteit Leuven, Belgium Jonas BerggrenВ©Folio DonвЂ™t Miss The New International Myeloma Guidelines New Guidelines recommend that serum free light chain (FLC) assays be used in the initial evaluation of suspected myeloma1 Guidelines published by the International Myeloma Working Group recommend Serum FLC for use in screening, at diagnosis, for prognosis and when monitoring patients with multiple myeloma and related disorders. is a simple blood test to quantitate free kappa (Оє) and free lambda (О») immunoglobulin light chains. Email: [email protected] for your free copy of the new myeloma guidelines 1. Dispenzieri et al. Leukemia 2009:23:215-224 FreeliteTM & HevyliteTM are trademarks of The Binding Site Group Ltd, Birmingham, UK. THE BINDING SITE GROUP LTD United Kingdom Tel: +44 (0)121 436 1000 Fax: +44 (0)121 430 7061 www.freelite.co.uk www.cli-online.com & search 24724 Go to: www.freelite.co.uk to contact us in: UK, USA, Germany, Austria, France, Spain, Canada and 70 other countries COMING SOON! See what the HevyliteTM ratio offers you. Visit: www.freelite.co.uk for more information. вЂ“ Issue NВ°5 вЂ“ October 2009 20 Microbiology Rapid screening for antibiotic-resistant bacteria As bacterial resistance to antibacterial agents continues to spread globally, the role of infection control is increasingly important in the battle against multidrug-resistant hospital acquired infections. New developments in chromogenic culture media help clinical laboratories to screen patient samples quickly and easily for the presence of meticillin-resistant Staphylococcus aureus, Extended Spectrum Beta-Lactamase-producing organisms and Vancomycin Resistant Enterococci, allowing infections caused by these multidrug-resistant bacteria to be identified rapidly so that infection control procedures can be initiated at the earliest opportunity. by J. E. C. Beaves Microbiologists and infection control professionals worldwide are increasingly concerned about the growing number of clinically significant microorganisms that have developed resistance to commonly prescribed antibiotics. Such strains are a common cause of hospital-acquired infections (HAI). More and more frequently, they limit the choice of therapy available (often resulting in the empiric use of more expensive antimicrobial agents), cause increased morbidity and mortality in patients, and result in prolonged hospital stays. Measures to control such infections and to provide better outcomes are being introduced throughout the developed world. Meticillin-resistant Staphylococcus aureus infection was, for many years, the most notorious of these HAIs, but now other, equally serious threats are emerging. Health professionals worldwide are showing particular concern about the increasing prevalence of Extended Spectrum Гџ-Lactamase (ESBL)-producing organisms and Vancomycin Resistant Enterococci (VRE). Anxiety about these emerging problems is now spreading to the general public as new вЂњsuper bugsвЂќ increasingly feature in the news headlines. A joint working group from the European Centre for Disease Prevention and Control (ECDC) and the European Medicines Agency (EMEA) recently produced a technical report in response to the widening gap between the increasing prevalence of multidrug-resistant bacteria and the decline in development of antibacterial agents aimed at treating infections with these organisms. The report observed that вЂњemerging and increasing resistance has become a threat to public health in Europe and globally, so that we are now facing the possibility of a future without effective antibiotics for several types of bacteria that cause infection in humans.вЂќ Some of the most common multidrug-resistant bacteria isolated from blood cultures (and therefore with the potential to cause serious Table 1. Bacteria responsible for bloodstream infections and resistances used as markers for resistance to multiple antibiotics. infections) in Europe, as identified in the ECDC/ EMEA report, are shown in Table 1. The ECDC/EMEA working group reported that very few antibacterial agents with new mechanisms of action are under development to meet the challenge of multidrug-resistant bacteria and there is a particular lack of new agents to treat infections due to multidrug resistant Gram-negative bacteria, such as ESBL-producers. Consequently, rapid detection of these resistant organisms is increasingly important in order to initiate the most appropriate treatment and the necessary infection control procedures at the earliest opportunity. Recent advances in the development of chromogenic culture media allow MRSA, VRE and ESBL-producing microorganisms to be detected directly from clinical specimens in just 18-24 hours, providing an important rapid screening tool for use in the fight against multidrug-resistant bacteria. MRSA MRSA is the most common cause of antibiotic-resistant HAI in many parts of the world, including Europe, the Americas, North Africa, the Middle East and the Far East. In Europe, 31 countries participating in the European Antimicrobial Resistance Surveillance System (EARSS) reported 31,591 invasive S. aureus isolates in 2007, 22% of which were MRSA. The proportion of MRSA isolates varied across Europe, from 3% or less in northern countries to greater than 50% in some southern countries. However, the most recent figures show that more European countries are reporting decreasing trends in MRSA rates compared with previous years, indicating that improved control efforts can have a positive effect on MRSA levels in hospitals, even in high endemic countries. In England, for example, there has been mandatory reporting of MRSA bacteraemias since 2005. Since that time, intense scrutiny of the practices of individual hospitals, improved antibiotic prescribing policies and infection control practices have assisted in reducing the number of cases reported to the Health Protection Agency (HPA) to more than half - with 2932 cases reported in the financial year 2008/2009 compared to 7096 in 2005/2006 [Figure 1]. 21 вЂ“ Issue NВ°5 вЂ“ October 2009 Figure 1. MRSA bacteraemias reported in English NHS acute trusts. The importance of screening as part of an effective infection control programme to limit the spread of MRSA is well recognised and compulsory MRSA screening in elective admissions was introduced in England in April of this year. The effectiveness of a screening programme relies on the speed and accuracy of results so that colonised patients can be quickly and reliably targeted for isolation, decolonisation and appropriate treatment. Improved MRSA detection Chromogenic MRSA culture media allow the presumptive identification of MRSA to be achieved from patient swabs or clinical isolates. Oxoid Brilliance MRSA Agar has been shown to be one of the most selective MRSA chromogenic media available and can provide reliable results in as little as 18 hours. This medium detects the phosphatase activity of MRSA using a novel chromogenic compound. When cleaved, the chromogen produces distinctive denim blue colonies. Antibacterial agents within the medium inhibit the growth of competing organisms, including MSSA, while additional compounds suppress the expression of phosphatase activity in other staphylococci. These properties result in a highly selective medium that demonstrates excellent sensitivity and specificity and allows results to be obtained up to six hours earlier than alternative chromogenic media. The medium demonstrates exceptional positive and negative predictive values (98.1% and 99.2% respectively). These are higher than those claimed by many PCR-based systems and yet are achieved at a fraction of the cost. In circumstances where patient isolation facilities are in short supply, leading to the cohorting of MRSApositive patients, good positive and negative predictive values are essential. False-positive results could lead to MRSA-negative patients being put at increased risk of infection by prolonged stays in MRSA cohorted wards, in addition to the unnecessary and improper use of вЂ�last resortвЂ™ antibiotics. False-negative results could prevent an MRSA-positive patient from being isolated and receiving appropriate treatment. In an additional comparative trial, it was concluded that the excellent selectivity of Oxoid Brilliance MRSA Agar required fewer confirmatory tests, concluding that it is a reliable and economical option for clinical laboratories. ESBL-producing micro-organisms Extended Spectrum Гџ-Lactamase (ESBL)producing microorganisms are another class of resistant bacteria that have been on the rise since they were first characterised in Europe in 1983. The relevant enzymes are found in a family of organisms known as the Enterobacteriaceae, which includes common commensal flora, such as Klebsiella pneumoniae and Escherichia coli. Enterobacteriaceae are a significant cause of nosocomial and community-acquired infections. Due to consistent reporting over the last 26 years, it has been possible to identify a number of factors that have contributed to the increase of these infections, including the overuse of antibiotics in humans and animals, hospital cross-infection, human migration and changes in the food chain [1]. The main therapeutic options for the treatment of Enterobacteriaceae infections are Гџ-lactam antibiotics (broad spectrum penicillins and cephalosporins). However, ESBLs confer resistance to these compounds. Furthermore, ESBL resistance genes are encoded on freely transmissible genetic elements, greatly increasing the risk of spread of resistance to other organisms. The European Antibiotic Resistance Surveillance System has been monitoring trends in the numbers of bacteria producing these enzymes since 2000. Previously, ESBLs were mostly found in Klebsiella species, with infections restricted to certain vulnerable patient groups, often in intensive care. However, a new class of ESBL (referred to as CTX-M enzymes) has emerged, which is widely detected among E. coli and is most frequently found in hospital- and community-acquired urinary tract infections. www.cli-online.com & search 24778 вЂ“ Issue NВ°5 вЂ“ October 2009 22 Microbiology are confined to Enterococcus faecium, although a few cases have been confirmed as originating in Enterococcus faecalis. Typing has confirmed that there are distinct differences between the subpopulations that are found in hospitals, and human commensal and animal strains. The hospital-acquired strains have additional genomic content with associated virulent factors (and ampicillin resistance in European strains). Resistance to ampicillin tends to precede an increased VRE rate, which emerges within several years [3]. The latest EARSS report (2006) showed that there has been a continuous increase in the number of invasive E. coli and K. pneumoniae isolates that are resistant to third generation cephalosporins and contain the ESBL enzymes. The report included data from 800 laboratories in 31 countries and showed a higher than 10% occurrence in half of the enrolled countries [1]. As with MRSA, there is a definite split between the northern and southern European countries. The occurrence of ESBL isolates is still considered low in the most northern European countries compared to southern and eastern countries. The lowest occurrence of ESBL clinical isolates is in the Nordic countries. However, even there, awareness has heightened recently as an outbreak of ESBL-producing Klebsiella has been reported amongst newborns and children in a Swedish hospital. Although the outbreak has been relatively small (to date; four children infected and an additional four identified as carriers), the effects have been devastating, claiming the lives of three newborn babies. In southern and eastern Europe, there is an increasingly high occurrence of ESBLs in both nosocomial and community settings [1]. . ESBL-producing isolates are normally resistant to a number of different antibiotic families (including beta-lactams, fluoroquinolones, aminoglycosides and trimetoprim-sulfametoxazole), which severely limits treatment options and increases the multidrug-resistant ESBL strains in both medical and social settings. Rapid detection of ESBL production is, therefore, extremely important, so that an effective antibiotic can be prescribed. Rapid detection of ESBL-production A new chromogenic medium, Oxoid Brilliance ESBL Agar, has been developed for the presumptive identification of ESBL-producing micro-organisms direct from clinical samples, in just 24 hours. This carefully formulated medium distinguishes between ESBL-producing E. coli and ESBL production in the KESC group of bacteria (Klebsiella, Enterobacter, Serratia and Citrobacter). Differentiation is achieved by using two chromogenic compounds which target galactosidase and glucuronidase activity. The KESC group of organisms produce galactosidase, resulting in green colonies on the agar, while E. coli express galactosidase and glucuronidase, producing easily distinguished blue colonies [Figure 2]. Occasionally, E. coli will be galactosidase negative, but these organisms produce pink colonies. Proteus, Morganella and Providencia do not utilise either chromogen, but produce tancoloured colonies with a brown halo due to the Figure 2. ESBL-producing E. coli (blue colonies) and KESC organisms (green colonies) growing on Brilliance ESBL Agar. deamination of tryptophan. The semiopaque background of the medium contrasts with the brightly coloured colonies and allows clear and easy identification of target organisms. The medium contains cefpodoxime and additional antibacterial agents to inhibit non-ESBL-producing Enterobacteriaceae and to suppress the growth of less resistant AmpC organisms and other non-ESBL flora, including Stenotrophomonas maltophilia. Inhibition of these organisms reduces the incidence of false-positive results compared to traditional culture media, minimising the need for confirmatory testing. An evaluation of the medium was performed using a variety of isolates, including CTX-M, TEM, SHV and K1-hyper-producing strains, from clinical* and other sources. Results indicated that K1-hyper-producing (non-ESBL) strains were inhibited while all representative ESBL strains grew. The agar was selected by MOSAR (Mastering Hospital Antimicrobial Resistance in Europe) for use in a pioneering European ESBL prevalence study (for further information visit www.mosar-sic.org). Tracking VRE rates has become even more important now that a link between ampicillin resistance and VRE rates in hospital isolates has become apparent. Within Europe, there is much variability between surveillance systems that have been set up to collect data on vancomycin resistance in enterococci, and in some countries no data is collected. Due to these missing and variable data, it is difficult to perform reliable statistical analysis. However, the EARSS report for 2007 suggests that some trends have emerged. There have been increasing VRE rates in some countries, including Ireland, Germany and Greece, whereas in the Nordic countries and the Netherlands there is low VRE prevalence. Austria, Portugal and Italy have seen decreases, but there is insufficient evidence to link this to measures that these countries took to contain outbreaks [3]. VRE-related infections are usually seen in hospital patients who are already very ill, in particular those who are immunocompromised, those who have had previous treatment with certain antibiotics (particularly cephalosporins and glycopeptides), those on a prolonged hospital stay, or those in specialist units, such as intensive care, oncology or renal units. Prompt identification of infection in these vulnerable patient groups is extremely important in order to improve outcomes and to prevent the spread of infection to other patients. Vancomycin Resistant Enterococci (VRE) Glycopeptide resistance in enterococcal species is also increasing and, in particular, Vancomycin Resistant Enterococci (VRE) have emerged as significant nosocomial pathogens. This is thought to be due to the increased use of vancomycin for treatment of MRSA, particularly in the United States, and the use of a vancomycin-like glycopeptide (avoparcin) as a growth promoter in animal husbandry in Europe [2]. There are a number of vancomycin resistance mechanisms, of which the most genetically mobile and widespread are VanA and the less common VanB. The first report of VRE occurred in the early 1980s in Europe. Most VRE strains Figure 3. E. faecium (purple) and E. faecalis (light blue) colonies growing on Brilliance VRE Agar. 23 Enhanced detection of VRE Recently, Oxoid Brilliance VRE agar, a new chromogenic screening plate for the detection of VRE, has become available. This medium provides presumptive identification of vancomycin resistant Enterococcus faecium and Enterococcus faecalis, from a faecal sample, swab, isolate or suspension, within just 24 hours. Differentiation between the two species is achieved using two chromogenic compounds - one that targets phosphatase activity (present in both species) and another that targets О±-galactosidase activity (present in E. faecium). This results in light blue E. faecalis colonies, which are easily distinguished from the indigo/purple colonies of E. faecium [Figure 3]. help clinicians to provide the most effective care, but can contribute to more widespread epidemiological studies and to our understanding of these important pathogens. References 1. Coque TM, Baquero F, Canton R. Increasing Prevalence of ESBL-producing Enterobacteriaceae in Europe. Eurosurveillance 2008; 13:47. 2. Bell JM, Paton JC, Turnidge J. Emergence of Vancomycin Resistant Enteroccocci in Australia: Phenotypic and Genotypic Characteristic of Isolates. J. Clin. Microbiol 1998; 36, 2187-2190. 3. Werner G, Coque TM et al. Emergence and spread of Vancomycin Resistance Among Enterococci in Europe. Eurosurveillance 2008; 13:47. A full list of references is availble from the author. * Clinical isolates were provided by Dr. Maurine A. Leverstein-van-Hall, Clinical Microbiologist, University Medical Centre Utrecht (UMCU)/National вЂ“ Issue NВ°5 вЂ“ October 2009 Institute for Public Health and Environment (RIVM), Netherlands, and Professor Youri Glupczynski, University Clinic of the Catholic University of Louvain (UCL) Mont-Godinne, Belgium. The author James E. C. Beaves BSc.(Hons) Clinical Applications Manager Oxoid, Thermo Fisher Scientific Basingstoke, Hants, UK. Email: [email protected] www.cli-online.com & search 24796 Shaping the future of Enzyme Extraction The growth of competing flora, including E. gallinarum and E. casseliflavus (both of which are intrinsically resistant to vancomycin) is suppressed by the inclusion of vancomycin and additional antibiotics in the medium. The medium was evaluated in a clinical trial, using a panel of 120 clinical isolates, and demonstrated a sensitivity of 94.7% and 100% at 24 and 48 hours respectively. Routine screening Unlike some other rapid methods for the identification of resistant organisms, which require expensive or specialised equipment, chromogenic culture media can be adopted easily by clinical laboratories of any size. Brilliance MRSA, VRE and ESBL media are supplied in pre-poured agar plates that are ready to inoculate, and give easyto-read results within 24 hours or less, directly from clinical samples. This speed, convenience and ease of use make chromogenic media a valuable option for routine screening for significant resistant microorganisms by clinical laboratories. As resistance mechanisms, such as those described above, continue to challenge health professionals around the world, it is important to have such reliable and easily accessible tools for local screening programmes, which not only New Custom Matching Service from BBI Enzymes вЂў Clinical Chemistry вЂў Blood Glucose Monitoring вЂў Urine Test Strips Our new custom specification service is tailor made to your exact requirements for Horseradish Peroxidase and Glucose Oxidase, promising highly competitive pricing, batch to batch consistency on large volumes, and fast turnaround. 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Contact Ian Southam on +44 (0) 1495 790 678 or email [email protected] Visit BBI Enzymes at Medica 2009 Hall 1, stand F24 BBI Enzymes, Blaenavon, Gwent, NP4 9RL, UK T: +44 (0) 1495 790678 E: [email protected] your no.1 choice for natural enzymes www.bbienzymes.com www.cli-online.com & search 24822 вЂ“ Issue NВ°5 вЂ“ October 2009 24 Haematology Case study: how a general hospital lab improved its blood film review procedures The haematology lab of Middlemore Hospital, South Auckland, New Zealand, had been using the traditional light microscope to review blood films. The laboratory is now using an automatic morphology system to review the majority of its films. We spoke to John Peters, Scientist in Charge of the haematology laboratory at Middlemore hospital, to see how the system performs in practice compared with the manual system. Q: Could you give us some background regarding the hospital at Middlemore and the main activities of the haematology lab? A: The hospital has approximately 900 beds at its main facility but also has two smaller satellite facilities a short distance away. The hospital is a general hospital and provides mental health services, general surgery, paediatrics, womensвЂ™ health and geriatics. It also houses the largest maternity unit and the largest accident and emergency unit in New Zealand, as well as the national burns unit and the largest renal inpatient and outpatient unit in the country. The satellite sites provide extensive outpatient services. The laboratory serves all these areas and also supports a haematology ward and clinic that deal with chronic haematological patients, but not with acute leukaemia or oncology patients. The laboratory has four consultant haematologists covering both the lab, haematology ward and clinics. There is one laboratory registrar and approximately 18 full time equivalent staff, 12 of whom are scientists and the others of whom are technicians. Q: What patient population does your hospital/ lab serve and how many blood films does the lab receive per year? A: Middlemore hospital is one of three large general hospitals serving the City of Auckland, which has a population of 1.4 million people. Middlemore hospital serves the southern section of the greater Auckland area with a population that is currently approximately 480,000 people but which is growing faster than that in any other area of New Zealand. The haematology lab processes about 180,000 full blood counts per year and of these, reviews approximately 46,000 films. The population served is the poorest in the country and there are many abnormal results. Q: Since when has the lab been using automated image analysis? A: We have been using the CellaVision DM96 since May 2009. The lab also changed its cell counters at that time and currently uses Sysmex XE5000 Alpha systems. Q: How do you find the DM96 system compared with the manual morphology system? A: There are two main advantages that we have identified so far. These are speed and the fact that the work is physically easier on the staff. We have found the review rate to be almost twice as fast as manual reviewing when experience staff are using the system. This is a big advantage for us, as all our staff are experienced with the exception of students undergoing training, and interns. In the past we have had a few problems with staff suffering from over-use of a combination of microscopes and computers. The DM96 has reduced these problems substantially since there is less use of the manual microscope and the actions necessary to focus and move the stage. Approximately 4% of films are still reviewed manually and the majority of these are paediatric samples. Having to look at a fixed number of cells defines an end point to the review, and therefore an end to the process. The excellent WBC categorisation and excellent morphological appearance of the cells makes the review much easier and more accurate. We are much more aware of abnormal cells and they cannot be overlooked. If the DM96 identifies a blast cell, for example, the staff member must look at this and reclassify it if necessary. We have the remote viewing stations set up with two flat screen monitors running off a single PC, John Peters, Scientist In Charge of Middlemore Hospital Haematology Laboratory. making the film reviews a lot less labour intensive and physically less demanding than manual microscopic reviews. One monitor displays the DM96 images and the other displays the lab computer results and analyser scatter plots. Images that we require for presentation work and for our library of abnormal images are kept either on CD or flash drive. The DM96 is extremely good for auditing purposes as the scientist/technician who has reviewed the images can easily be identified from the log in on the computer. If a user is mis-categorising cells on a regular basis then that user can be identified. The DM96 is an excellent instrument for teaching new staff members RBC morphology, WBC morphology and platelets morphology, and it has an excellent library of images on board that can be augmented. We donвЂ™t use the RBC morphology recognition system but categorise the morphology ourselves and this is both fast and accurate. Q: Are there any improvements that you would like to see in the system? A: We would welcome a means of looking at the tail of the film for platelet clumps, microfilaria and clumps of abnormal cells. We would like to see a modification of the DM96 main analyser so that it could detect the last film in a cassette and then reject it quickly and move on to the next one. This would speed up the processing of samples a lot when only partially loaded cassettes are introduced into the instrument. Q: Finally, could you summarise how the installation of the DM96 in your lab has helped both clinicians and their patients? A: The clinicians donвЂ™t use the DM96 but the instrument allows the lab to analyse and report on the samples in a more timely fashion and in a more standardised and accurate manner. Education of the clinical staff is also a lot easier with easy access to a large variety of cells from the comprehensive library and from patient samples. CellaVisionвЂ™s new analyser, the DM1200, is now available, see page 35. "We are the osmometer people." 30 years of success: our company has matured! To be honest on this, our 30th anniversary, we are rather proud of what we have achieved during the past three decades. Thirty years in which we grew together as a team, but also thirty years in which, in a sometimes difficult and always highly competitive market, we asserted ourselves due to our knowledge, our high-quality products and вЂ“ last, but not least вЂ“ due to a trusted cooperation with our customers, to whom we would here like to express our thanks. We have also utilised our strengths in the development of a new instrument, the Chloridmeter CM20: our ability to produce high quality products and our knowledge of what will satisfy the demands of our customers. These are strengths that years ago resulted in our companyвЂ™s philosophy: вЂњWe are the osmometer people.вЂњ And today we are glad to be able to add: вЂќAlways were, always will be.вЂќ We are not an aloof large organisation: we know our customers personally, are familiar with their work and their problems, and thus know what they require from us and the quality they expect from our products. We have remained a medium-sized, private company for 30 years, but we have also become a global player with customers in more than 60 countries. In its traditional casing! Now, in 2009, our anniversary year, we have moved from Berlin- SchГ¶neberg after many years to our new premises in Berlin- Charlottenburg. A lot of space for new ideas! www.cli-online.com & search 24777 Ana l yt e of t he m ont h вЂ“ Issue NВ°5 NВ°6 вЂ“ October 2009 2008 26 Focus on Finland Novel biochemical markers of bone remodelling in the management of osteoporosis Currently available bone turnover markers are extensively used in research applications and have also shown clinically interesting associations. Recent advances in bone cell biology have led to the development of potential new markers. Novel candidates include osteoclastic enzymes, non-collagenous proteins, markers of bone matrix properties and regulators of bone cell activity. by Dr Kaisa K. Ivaska Bone is a metabolically active organ that is continuously subjected to resorption and formation on the surface of the trabecular bone area (typically in the interior of the bone) and in the Haversian canals. During bone resorption, multinucleated osteoclasts degrade existing bone matrix. Resorption is followed by bone formation, during which osteoblasts synthesise new bone matrix which is gradually mineralised. Most of the bone turnover takes place in trabecular bone due to its higher surface area. Quantitative changes in bone metabolism can be assessed easily and non-invasively by measuring biochemical markers of bone remodelling in serum or urine. These markers, also known as bone turnover markers, are skeletal proteins or their fragments, or enzymes released from bone cells during bone turnover. Formation and resorption are usually tightly coupled in time and space. Therefore, in disease states where both events are coupled and change in the same direction, such as in osteoporosis, any marker reflects the overall rate of bone turnover. Biomarkers for osteoporosis management Come to see our вЂў enlarged selection of infectious disease kits вЂў expanded neonatal screening panel in Medica Stand 3/C 83 Ani Labsystems Ltd. Oy [email protected] [email protected] www.cli-online.com & search 24768 The most widely used currently available markers include bone specific alkaline phosphatase (boneALP), osteocalcin (OC) and the N-terminal propeptide of type I collagen (PINP) for bone formation, and crosslinked C and N telopeptides (CTX and NTX) and deoxypyridinoline (DPD) for bone resorption. None of the current markers are used to diagnose osteoporosis. Instead, bone mineral density (BMD) measured by dual energy X-ray absorptiometry (DXA) is the single best method available for confirming the diagnosis of osteoporosis, defined as BMD value 2.5 standard deviations or more below the mean of the young female adult population. However, bone markers provide more dynamic and rapid measures of skeletal status. They have been investigated in osteoporosis for use in three main areas: (i) monitoring anabolic or anti-resorptive therapy, (ii) predicting bone loss and the risk of developing osteoporosis, and (iii) identifying individuals with high risk of fracture. Bone markers are useful tools in assessment of response to anti-resorptive or anabolic therapy. Significant changes in markers occur within months whereas it usually takes a few years to detect a significant change in BMD. Monitoring the efficacy of bone-active drugs is currently the most promising and widely known clinical application. Another area where markers may be of use is bone loss. Bone loss can be assessed by DXA if serial measurements are performed but it takes years to detect significant changes due to measurement imprecision and the relatively slow rate of change in BMD. The assessment of bone markers could be of use in identifying individuals who have increased bone turnover and who may be at the greatest risk for bone loss and of developing osteoporosis We have found that women who had constantly high bone turnover in serial measurements lost significantly more bone over five years than women with constantly low turnover. They also progressed to the level of osteoporosis more frequently [1]. Serial assessment of bone turnover may thus assist in decision-making and targeting preventive measures. One of the most important challenges in the management of osteoporosis is, however, to identify individuals who are at high risk for fragility fractures. Low BMD is a strong risk factor for fractures but there is a great overlap in BMD of individuals with and without fracture. Apparently, BMD is only one of a number of risk factors and can capture only one aspect of the likelihood of fracture. Due to the low sensitivity of BMD testing, there is a need for additional measures to identify individuals who are at high risk for fractures and who might need treatment, despite not reaching the diagnostic threshold for osteoporosis. During recent years risk-based assessment has been applied and the Fracture Risk Assessment Tool (FRAX) integrating clinical risk factors has been developed to evaluate fracture risk of patients. High levels of bone markers have been shown to be associated with fractures independently of BMD in postmenopausal women in several prospective studies [2]. The results have been most consistent with resorption markers. Markers could add information, particularly for individuals with normal or moderately low bone mass (osteopenia), who 27 Overview on existing bone turnover markers (black) and novel candidate markers (orange). Abbreviations: PINP, N-terminal propeptide of type I collagen; Dkk1, Dickkopf 1; boneALP, bone-specific alkaline phosphatase; OPG, osteoprotegerin; sRANKL, soluble receptor activator of nuclear factor kappa B; OC, osteocalcin; CTX and NTX, C- and N-terminal crosslinked telopeptide of type I collagen; DPD, deoxypyridoline; TRACP5b, tartrateresistant acid phosphatase 5b. are asymptomatic but have increased bone metabolism and may thus be at increased risk for developing osteoporosis and osteoporotic fractures in the future [3]. Novel candidate markers of bone remodelling Currently available bone turnover markers are extensively used in research applications and have shown clinically interesting associations, but they also have some limitations. Markers reflect quantitative changes but do not provide information on structural abnormalities of bone. Furthermore the relative contribution of trabecular, cortical and periosteal bone may vary depending on disease, age or therapy. Some analytes also have high variability or are not bonespecific. Recent progress in the identification of important pathways in bone physiology has led to the development of potential new biochemical markers. Candidates include osteoclastic enzymes, non-collagenous proteins and markers of bone matrix properties. In addition, regulators of bone cell activity have been studied. fractures, in particular for vertebral fractures [5]. Cathepsin K is a member of the cysteine protease family and is secreted into the resorption lacuna, where it contributes to the cleavage of collagen molecules and leads to degradation of the organic matrix. Cathepsin K is endocytosed with the degradation products, transported through the cell and released into the circulation where it can be detected by immunoassay. It is a potential marker of osteoclast number but because the circulating concentrations are low, the accurate determination with current assays remains challenging. Non-collagenous matrix proteins and their fragments Although circulating osteocalcin is widely used as an index of bone formation, some of the fragments are derived from the bone resorption process when osteocalcin is released from the bone matrix. These fragments are rapidly cleared through glomerular filtration and accumulate in the urine. Urinary OC fragments вЂ“ Issue NВ°5 вЂ“ October 2009 consist of the mid-molecule portion, which appears to be protected from degradation in the kidney. Immunoassays for urinary osteocalcin fragments have been developed and theoretically, urinary osteocalcin fragments may constitute a more specific bone resorption marker than type I collagen-related fragments [6]. Other potential non-collagenous markers include bone sialoprotein, for example. It has been shown to be a product of active osteoblast, but is strongly upregulated in a variety of human primary cancers, including breast, prostate and lung cancer. The limited clinical data suggest that BSP may be interesting for use in patients with malignant bone disease. Markers of bone matrix properties Pentosidine is formed by non-enzymatic glycation of type I collagen when sugar is present in extracellular matrix. It is a crosslinking molecule although the precise location in collagen helix is not known. High pentosidine content in bone matrix has been associated with decreased resistance to fracture. High serum pentosidine has recently been shown to be moderately associated with vertebral fracture in postmenopausal women with type 2 diabetes [7]. C-terminal crosslinked telopeptide of type I collagen, CTX, is a collagen fragment of eight amino acids, which exists in a nonisomerised and an isomerised form referred to as О±-CTX and ОІ-CTX, respectively. ОІ-CTX contains a ОІ-aspartyl peptide bond in L-enantiomeric form, which is considered to result from the ageing of extracellular proteins. The isomerisation process reaches a maximum of approximately three years after the bone is mineralised, and therefore the released ОІ-CTX reflects the degradation of relatively old bone. On the contrary, the non-isomerised О±-CTX presumably results from the degradation of relatively young bone. The urinary ratio of these Osteoclastic enzymes Tartrate-resistant acid phosphatase 5b (TRACP5b) belongs to the family of five isoenzymes of acid phosphatases. Two isoforms of TRACP exist in circulation. TRACP5a originates from macrophages and dendritic cells, whereas TRACP5b is derived from the osteoclasts. TRACP5b is believed to destroy the endocytosed bone matrix degradation products during transcytosis through the osteoclast. TRACP5b is highly expressed and secreted by both resorbing and non-resorbing osteoclasts into the circulation. Recent data show that it reflects the number of osteoclasts rather than osteoclastic activity. S-TRACP has very little circadian variation and is not influenced by renal function or by food intake [4]. Using a population-based design, serum TRACP5b was found to be predictive for forthcoming All you need for your assay development HyTest Ltd produces and delivers high-quality immunological reagents for industrial and research applications all around the world. Focusing on providing good value for its customers, HyTest strives uncompromisingly for a comprehensive understanding of the constantly developing needs of industrial and research communities. This is the HyTest formula for success. HyTest Ltd., Intelligate, 6th floor Joukahaisenkatu 6, FI-20520 Turku, Finland Phone: +358 2 512 0900. Fax: +358 2 512 0909 www.hytest.fi [email protected] www.cli-online.com & search 24787 вЂ“ Issue NВ°5 NВ°6 вЂ“ October 2009 2008 28 two forms can be measured by immunoassays and it gives information on collagen maturation which could contribute to bone matrix quality. This is supported by studies which have demonstrated that the urinary О±/ОІ CTX ratio is predictive for fractures independent of BMD and bone turnover, suggesting that the ratio provides additional information beyond bone mineral density and turnover rate [8]. Regulators of bone cell activity Osteoprotegerin (OPG) and receptor activator for nuclear factor kappa B ligand (membrane-bound RANKL, secreted sRANKL) are expressed by osteoblasts and are important local regulators of bone turnover. The interaction of RANKL with the receptor on osteoclast precursors induces the differentiation and maturation of osteoclasts, while OPG acts as a decoy receptor and prevents osteoclastogenesis. At present it remains unclear what proportion of circulating OPG is monomeric, dimeric or bound to RANKL and which form is the most biologically relevant to measure. Due to these limitations, conflicting data exist. It is also unclear how systemic concentrations reflect local cytokine concentrations in bone. Wnt signalling, on the other hand, plays a key role in the regulation of osteoblastic activity. Different secreted proteins inhibit Wnt signalling in osteoblasts, such as soluble frizzled-related proteins (sFRPs), Wnt inhibitory factor-1 (WIF1), Dickkopfs 1-4 (Dkk1-4) and sclerostin. An assay for circulating Dkk-1 has been developed and increased levels have been detected e.g. in patients with breast cancer bone metastases [9]. The role in postmenopausal osteoporosis remains to be determined. With the advances in bone cell biology, other regulatory molecules are also emerging. Recent findings suggest that bone is an important endocrine organ producing Focus on Finland hormones, such as FGF-23 regulating phosphate metabolism in the kidneys and uncarboxylated osteocalcin affecting adipose tissue and pancreatic beta cells [10]. Conclusions Progress in the characterisation of important biological pathways in bone cell biology and the components of bone matrix has led to the development of new candidate markers. Of these, TRACP5b, the О±/ОІ CTX ratio, serum pentosidine and urinary osteocalcin fragments have been shown to be associated with fracture risk in preliminary clinical studies. Post-translational modifications of bone matrix proteins are interesting candidates since they may have a role in determining the mechanical competence of bone. Thus they would complement the existing markers by providing information on qualitative changes in bone matrix properties. Further studies are required to elucidate the clinical value of novel markers in osteoporosis, alone and in combination with existing markers. The list of available markers should further expand in the near future. References 1. Ivaska KK, Lenora J, Gerdhem P, Akesson K, Vaananen HK, Obrant KJ. Serial assessment of serum bone metabolism markers identifies women with the highest rate of bone loss and osteoporosis risk. J Clin Endocrinol Metab 2008; 93(7):2622-32. 2. Garnero P. Markers of bone turnover for the prediction of fracture risk. Osteoporos Int 2000; 11 Suppl 6:S55-65. 3. Sornay-Rendu E, Munoz F, Garnero P, Duboeuf F, Delmas PD. Identification of osteopenic women at high risk of fracture: the OFELY study. J Bone Miner Res 2005; 20(10):1813-9. 4. Halleen JM, Tiitinen SL, Ylipahkala H, Fagerlund KM, Vaananen HK. Tartrate-resistant acid phosphatase 5b (TRACP 5b) as a marker of bone resorption. Clin Lab 2006; 52(9-10):499-509. 5. Gerdhem P, Ivaska KK, Alatalo SL, Halleen JM, Hellman J, Isaksson A, Pettersson K, Vaananen HK, Akesson K, Obrant KJ. Biochemical markers of bone metabolism and prediction of fracture in elderly women. J Bone Miner Res 2004; 19(3):38693. 6. I vaska KK, Kakonen SM, Gerdhem P, Obrant KJ, Pettersson K, Vaananen HK. Urinary osteocalcin as a marker of bone metabolism. Clin Chem 2005; 51(3):618-28. 7. Yamamoto M, Yamaguchi T, Yamauchi M, Yano S, Sugimoto T. Serum pentosidine levels are positively associated with the presence of vertebral fractures in postmenopausal women with type 2 diabetes. J Clin Endocrinol Metab 2008; 93(3):1013-9. 8. G arnero P, Cloos P, Sornay-Rendu E, Qvist P, Delmas PD. Type I collagen racemization and isomerization and the risk of fracture in postmenopausal women: the OFELY prospective study. J Bone Miner Res 2002; 17(5):826-33. 9. Voorzanger-Rousselot N, Goehrig D, Journe F, Doriath V, Body JJ, Clezardin P, Garnero P. Increased Dickkopf-1 expression in breast cancer bone metastases. Br J Cancer 2007; 97(7):964-70. 10. Fukumoto S, Martin TJ. Bone as an endocrine organ. Trends Endocrinol Metab 2009; 20(5):230-6. The author Dr Kaisa K. Ivaska University of Turku, Institute of Biomedicine Department of Cell Biology and Anatomy FI-20520 Turku, Finland and Clinical and Molecular Osteoporosis Research Unit Lund University Department of Orthopaedics MalmГ¶ University Hospital SE-20502 MalmГ¶ , Sweden Decreased bone mineral density in adults born with very low birth weight In a study published in the open-access medical journal, PLoS Medicine, Petteri Hovi and colleagues from the National Institute for Health and Welfare Helsinki, Finland evaluated skeletal health in 144 adults (ages ranging from 18 to 27 years) who were born preterm with very low birth weight. They showed that as adults these individuals have significantly lower bone mineral density than do their term-born peers and suggest that this finding translates into increased risk for osteoporosis in adulthood for these individuals. http://www.plosmedicine.org/article/ info%3Adoi%2F10.1371%2Fjournal.pmed.1000135 www.cli-online.com & search 24791 29 The Finnish diagnostics industry The Finnish diagnostics industry is rich with companies and research groups, which provide technologies and products to global markets. The diagnostics industry can be categorised into several speciality areas. These include companies that supply reagents and services, those supplying complete diagnostic kits, as well as companies that produce either instrumentation or electronic and mechanical components for incorporation into instrumentation in many different areas. The activities of some of these companies, as well as some of their new products, are outlined. Ani Labsystems Ltd This established Finnish diagnostics company is dedicated to the production and sales of immunoassays. The core business is high quality EIAs and FEIAs for infectious diseases and neonatal screening. For 25 years, and through changing ownership (formerly Labsystems), Ani Labsystems has been a key pioneer in the development of the first microplate arrays, screening of metabolic disorders for neonates and avidity methods. The company strongly invests in research and development, and state of the art production. The quality system complies with ISO 13485:2003 and IVD directive 98/79/ EC. The assays to diagnose infectious diseases include kits for antibodies to Mycoplasma pneumoniae, Chlamydia pneumoniae, Bordetella pertussis and Chlamydia trachomatis, as well as a full Toxo panel. The company offers PKU, TSH, Toxo-IgM, G6PD, 17-OHP, Galactosaemia and IRT for neonatal screening, and HIV and hepatitis B for blood screening. A unique assay to detect coeliac disease and a kit for quantitative IgD EIA are also available. HyTest Ltd HyTest Ltd is a biotechnology-oriented company based in Turku, that manufactures and markets high-quality immunological reagents for industrial and research applications. The product portfolio includes cardiac markers, hormones and toxins, human proteins, neuroscience-related reagents, infectious and autoimmune disease reagents. The company was established in 1994 and has experienced rapid growth over the subsequent years. The solid scientific background of its staff and continuous investments in R&D together with dedication to customer service and focus on global operations from day one have combined to ensure its success. HyTest is a leading supplier of several reagents worldwide, such as antibodies and antigens of troponin I and troponin complex. The inherent quality of its reagents and extensive customer support has helped the company to become a recognised name in the industry. The diagnostic industry is in a state of constant development. In response to this challenge, the company is continuously working to bring new products to the market as well as to improve the existing ones. Its R&D specialists maintain close ties with the scientific community and engage in joint research with Finnish and international academic diagnostic research organisations. HyTest is an ISO 9001:2000 certified company for the manufacture and distribution of immunological reagents. It strives to completely understand and satisfy the needs of its customers in both the industrial and research communities. Labquality Based in Helsinki, Labquality is an independent and impartial organisation founded in 1971 and owned by the Finnish Society of Clinical Chemistry, the Association of Finnish Local and Regional Authorities, 19 local hospital districts, the Finnish Medical Association, the Finnish Red Cross, the Association of Medical Service Providers and the Finnish Union of Experts in Science. Labquality provides External Quality Assessment (EQA) programmes for clinical laboratories and point-of-care tests performed in social and healthcare units. Its mission is to support, promote and improve quality in clinical testing, analytics and diagnostics. Labquality has 23 employees, and more than 100 experts are involved in the schemes. In 2009, Labquality had 4174 client laboratories based in 42 different countries. вЂ“ Issue NВ°5 вЂ“ October 2009 antibodies, as well as research and development, are conducted from the head office. Actim rapid tests are manufactured at the Joensuu site in Eastern Finland. The company employs around 100 highly trained professionals, and the product range covers monoclonal antibodies for a variety of markers. In order to meet the needs of different assay technologies, there are multiple clones for the most important analytes. MedixMAB monoclonal antibodies are manufactured in vitro using the latest manufacturing technology and serum or protein free culture media. All antibodies are fully characterised and purified by affinity chromatography. Diagnostic tests are marketed under the Actim range of rapid tests. Two of the most successful products are Actim PROM and Actim Partus. The Actim PROM rapid test reliably detects premature rupture of foetal membranes while Actim Partus is a fast and simple bedside test to estimate the ripeness of the cervix during pregnancy. In addition, the company has tests for respiratory (Actim Influenza A&B) and infectious diseases (Actim CRP) and gastroenterology (Actim Fecal Blood and Actim Pancreatitis). Reagena Ltd Based in Kauniainen, Reagena is an emerging diagnostics company in the Nordic region, focused on developing novel POC immunoassay tests in the areas of infectious disease testing and immunity. Aimed at developing biomarker-based novel rapid tests, ReagenaвЂ™s unique ReaMobile Diagnostic Test System commands respect in the POC testing market as it facilitates faster diagnosis and treatment of patients. As a product developed to address the needs of patients quickly and effectively, the wireless ReaMobile diagnostic test system has a unique place in the market as it fulfills the need for faster diagnosis in the immunoassay segment. With an ability to provide both quantitative and qualitative results, this test system utilising a mobile hand held reader is a valuable product addition. Labquality Medix Biochemica Medix Biochemica, owned by the Minerva Foundation, is a nonprofit foundation in medical research with half a century of extensive experience. The head office is located in Kauniainen, near Helsinki. Most activities, such as the production of monoclonal External Quality Assessment Services bioclin Controls, Calibrators and Reference materials for Internal Quality Assessment www.cli-online.com & search 24833 вЂ“ Issue NВ°5 вЂ“ October 2009 Product News 30 Tests for Toxoplasma gondii Ani Labsystems has updated the well-known Toxoplasma gondii IgG Avidity EIA test, which was the first Toxoplasma avidity test available globally. The new T. gondii mIgM EIA and IgG EIA tests are being developed to form a full вЂњToxo-packageвЂќ. The T. gondii mIgM EIA is a microcapture test, that can be run simultaneously with the T. gondii IgG EIA. The latter test provides quantitative results, and automatically generates the guidelines for the avidity assay (only 2 points dilution). Generally, the reliability of the original avidity test is now combined within a full package of tests that can be run fully automated as well as manually. For neonatal screening, a fluorometric Neonatal Toxoplasma gondii IgM FEIA is offered. The photometric version of the test, Neonatal Toxoplasma gondii IgM EIA, has recently been clinically evaluated; using over 1000 patient samples and almost 300 CDC samples, the test was found to have excellent specificity and sensitivity. useful for the development of high sensitivity cTnI assays, and also validated pairs suitable for lateral flow assays. A new generation of cTnI antibodies is currently under development. Work with brain natriuretic peptides is ongoing, and BNP, proBNP and NT-proBNP antibodies and antigens are available. In addition a new type of BNP/proBNP immunoassay, the вЂњsingle epitopeвЂќ sandwich assay, has been developed, which differs from the conventional format of sandwich immunoassays. In the single epitope sandwich assay, the capture antibody recognises the antigen (BNP or proBNP), whereas the detection antibody is specific to the complex of capture antibody and antigen. The single epitope sandwich approach has great advantages compared with conventional assays, especially in the case of unstable antigen detection. HyTest holds the intellectual property rights for this invention. Ani Labsystems Ltd Hytest Ltd Vantaa, Finland Turku, Finland Medica stand Hall 03/C83 Medica stand Hall 03/F44 detects influenza type A and B from respiratory samples. It has been proven that this test also detects the currently circulating Influenza A (H1N1) variant. Using this simple dipstick test, influenza can be diagnosed in just 15вЂ“20 minutes, including the time for sample collection. Similarly, the symptoms of respiratory syncytial virus (RSV), especially in infants, are very similar to the common cold and influenza. Diagnosing the disease early reduces the risk of serious complications, such as pneumonia. Actim RSV is a novel test that allows an instant RSV diagnosis to be made. All original and patented Actim tests are based on monoclonal antibodies developed and produced exclusively by Medix Biochemica. Medix Biochemica Kauniainen, Finland Medica stand Hall 03/F44 www.cli-online.com & search 24798 www.cli-online.com & search 24819 Troponin I and Brain Natriuretic Peptide cardiac markers Rapid diagnostic system www.cli-online.com & search 24803 HyTest specialists have been involved in cardiac troponin I (cTnI) studies for more than 15 years. Currently many well characterised antibodies directed to different regions of the cTnI molecule are available. Many of these antibodies are used in commercial assays. The company has determined antibody pairs and combinations Tests to differentiate between common cold, influenza and RSV Diagnosing viral infections is more important than ever. The influenza A (H1N1) pandemic has magnified the need for a reliable and fast method of testing symptomatic patients. Quick diagnosis is vital in managing the disease effectively with antiviral treatment. Actim Influenza A&B is a rapid qualitative in vitro assay that Action needed in pandemic scale. As the influenza A (H1N1) infection is sweeping the world, there is an urgent need for a reliable rapid test to diagnose patients with symptoms. www.actim.info www.cli-online.com & search 24788 www.medixbiochemica.com Copyright В© 2009 Medix Biochemica. We are also looking for new distributors for Actim tests! MA00066-01 The original Actim Influenza A&B rapid test is the answer you are looking for. The fast and reliable ReaMobile Rapid Diagnostic System is based on a novel lateral-flow immunoassay technology, which consists of a handheld ReaScan test reader, ReaScan rapid tests and ReaMobile software. The test procedure takes 10-15 minutes and requires minimal training. Results can be transferred wirelessly via a ReaLink Bluetooth device to a mobile phone, or via cable to a computer for data processing. The rapid test portfolio includes quantitative and qualitative applications for CRP, hsCRP, Strep A, and Puumala IgM among others. The test menu is steadily increasing to cover the important diagnostic markers analysed at the point-of-care. Licensing or OEM options are available. Reagena also manufactures a wide range of products for staining in microbiology, pathology and haematology. One of the most attractive and widely used products is REASTAIN Quick-Diff KIT, an optimised colour and fixative composition for the differential staining of cellular elements in peripheral blood smears. The staining procedure with readyto-use solutions takes only three minutes. REAGENA Toivala, Finland Medica stand Hall 03/F33 www.cli-online.com & search 24799 News in brief Severe stress can cause stroke Many patients urgently admitted to hospital with cerebral infarction state that they were under great stress over a prolonged period prior to suffering their stroke. This was shown in a unique patient study conducted in cooperation between the Sahlgrenska Academy at the University of Gothenburg and Sahlgrenska University Hospital, Sweden. The study is published in the scientific journal BMC Medicine. Nearly 600 patients were asked to complete a questionnaire in this study, no later than ten days after being admitted to Sahlgrenska University Hospital with acute cerebral infarction. In the questionnaire, the patients were asked to choose between six different alternatives to indicate how stressed they had felt before their stroke, from вЂњnever been stressedвЂќ to вЂњconstantly stressed over the past five yearsвЂќ. The patientsвЂ™ responses were compared with subjects in a healthy control group who were asked the same question. The study shows that there is a link to stress in those cases where the stroke is caused by atherosclerosis or to blood clots that have developed locally in the smaller vessels of the brain. The link was also found for those patients in whom it had not been possible to establish the cause of the stroke despite an extensive evaluation. On the other hand, the researchers could not see any independent correlation with stress for those patients who had had a stroke due to a blood clot from the heart. 31 вЂ“ Issue NВ°5 вЂ“ October 2009 Jane Norman, of the University of Edinburgh, and colleagues from NHS National Services in Scotland, researched a population-wide database of linked maternity records, infant health and death records in Scotland. They identified 1.49 million singleton births between 1980 and 2004, of which 90,000 were preterm births. Both spontaneous preterm births and medically-induced preterm births increased between 1980 and 2004; in absolute terms the rates of increase in each type of preterm birth were similar. Examining the database, the researchers found that maternal complications including pre-eclampsia and placenta previa played a decreasing role in preterm births over the period, whilst gestational and pre-existing diabetes played an increasing role. The researchers also found that there was an overall decline in stillbirth and in neonatal and perinatal deaths amongst preterm babies in the period covered вЂ“ although at 28 weeks gestation and beyond, stillbirths and perinatal deaths were reduced amongst medically-induced preterm babies, but were not reduced in babies who were spontaneously preterm. http://medicine.plosjournals.org http://www.biomedcentral.com Key role played by metalloprotease in survival of Leishmania parasite Leishmania is a deadly parasitic disease that affects over 12 million people worldwide, with more than 2 million new cases reported every year. Until recently, it was not known exactly how the parasite survives inside human cells. However, a team led by Dr Martin Olivier from the Research Institute of the McGill University Health Centre (RI-MUHC) and McGill University, Canada has elucidated the mechanism. It is hoped the new study, published in Science Signaling, will lead to development of the first prophylactic treatment for leishmania. The parasites enter the bloodstream and are ingested by macrophages where they block immune function and multiply, spreading to other tissues in the body. The researchers discovered that the metalloprotease GP63, which is found on the surface of the parasite, plays a role in neutralising the macrophagesвЂ™ defences. The GP63 protease directly activates other key molecules that negatively regulate the function of the host cell. The work is significant in that it is the first study that explains how the leishmania parasite blocks the immune function of macrophages. EARLY DIAGNOSIS OF ACUTE KIDNEY INJURY For more information please visit our booth K88 Hall 3 at MEDICA http://www.muhc.ca/media/news/ Spontaneous and medically induced preterm births contribute equally to the rising rate of preterm births Research published recently in the open access journal PLoS Medicine shows that the rising rate of preterm birth in Scotland is as much a result of an increase in spontaneous preterm birth as it is of preterm birth that is medically-induced to avoid risking the lives of the mother and child. The results emphasise that preterm birth, which remains the single biggest cause of infant death in many developed countries, continues to be a major obstetric and neonatal problem despite the reductions that there have been in stillbirths and perinatal deaths as a result of improved maternal medical care. Diagnostics BioPorto Diagnostics A/S Grusbakken 8 DK-2820 Gentofte Denmark Phone: Fax: E-mail: Web: (+45) 4529 0000 (+45) 4529 0001 [email protected] www.bioporto.com www.ngal.com www.cli-online.com & search 24783 вЂ“ Issue NВ°5 вЂ“ October 2009 News in brief 32 Breakthrough test can prevent fatal kidney failure Acute kidney injury has long been recognised as a devastating disorder, which often arises as a complication of other serious illnesses. Despite great efforts to understand and treat this condition the area has seen little or no improvement in over 60 years. Paradoxically, the incidence of this potentially deadly disorder has been rising during the last decades and is now reaching epidemic proportions. Each year, more than 13 million patients suffer acute kidney injury, of whom more than 30% have a fatal outcome. With a move that will prove to be a major advance in the diagnosis of this growing number of affected patients, Denmarkbased BioPorto Diagnostics has announced the successful development of a groundbreaking test called NGAL that can diagnose acute kidney injury. A test like this has been on the wish list of doctors worldwide for decades. Dr Lars Otto Uttenthal, CSO of BioPorto Diagnostics, said that the NGAL test had the potential to revolutionise the way kidney injury was managed. Existing methods of determining kidney injury do not provide useful information about the state of the patient until one to three days after an injury has occurred. In contrast, the NGAL test will reveal the occurrence Cal l Pos for ter 1 SHOW / 1 CONGRESS BIOLOGY WORKING FOR MEDICAL ADVANCE of kidney injury within a few hours. Dr Uttenthal said that doctors now had an opportunity to predict a patientвЂ™s outcome вЂ“ they could take proactive steps at an early stage that would prevent kidney injury from turning into the very dangerous state of established kidney failure. Thea Olesen, CEO of BioPorto, said that the icing on the cake was that the NGAL test was designed to be used on apparatus that already existed in laboratories and hospitals. In this way the company has ensured that the test will be affordable and readily available to doctors worldwide. In a market that is estimated to be worth over a billion dollars, the NGAL test could grab a considerable share, solidifying BioPortoвЂ™s position as the leading player in the NGAL market. The new NGAL test is developed in collaboration with one of the worldвЂ™s leading manufacturers of diagnostic tests and is currently being prepared for production. The test will be presented this month at ASN Renal Week in San Diego, USA, the yearly meeting point for kidney specialists from all over the world. www.bioporto.com Promising results for rapid viral diagnosis tests in emergency rooms Rapid viral diagnosis tests for respiratory diseases in children who arrive in emergency departments have the potential to reduce pressures on health systems by enabling doctors to reach a quicker diagnosis, according to Dr Quynh Doan of the British Columbia ChildrenвЂ™s Hospital in Vancouver, Canada, lead researcher of a recent study. Children who are admitted to emergency departments with cold and flu symptoms and fever undergo various diagnostic tests and are often prescribed antibiotics as a precautionary measure, even though viruses, which are often the cause, do not respond to antibiotics. The burden on health systems is huge, not only financially, but also in terms of the time and staff required to reach a diagnosis. Rapid viral diagnostic methods could help deliver fast, accurate diagnoses, and enable a much more appropriate use of antibiotics. The study included data from four trials, which together included 1,588 children. There was some evidence that rapid viral testing reduced use of other blood or urine tests, chest X-rays and antibiotics, but the results were not significant. However, the researchers suggest that further, sufficiently large studies could reveal the true impact of faster tests. esciencenews.com/topics/health.medicine High-sensitivity bone marrow aspiration technology enhances leukaemia cell detection INTERNATIONAL CONGRESS & EXHIBITION OF BIOLOGY 3*-4-5-6 NOV. 2009 вЂў CNIT Paris la DГ©fense вЂ“ France *Scientific conferences only GUEST OF HONOUR MOROCCO Organised by www.jib-sdbio.fr Superconducting Quantum Interference Device (SQUID) enhanced the ability to rapidly quantify the amount of nanoparticle-bound tumour cells in a sample at least 10 fold, and increased sensitivity of minimal residual disease measurements. Results of this proof-of-concept study are published in Cancer Research, a journal of the American Association for Cancer Research. These findings are a result of a collaborative research effort between Senior Scientific, LLC, and the University of New Mexico, USA. The study was funded by a small business innovation grant awarded by the National Cancer Institute, USA. The scientists developed the magnetic marrow biopsy needle in an effort to target tumour cells with nanoparticles and then preferentially extract the tumour cells with a magnetic needle. They used anti-CD34 antibody-loaded magnetic nanoparticles to detect CD34+ cells as an indicator of leukaemia. To quantify the cells recovered, they coupled this nanoparticle-mediated fishing for leukaemic cells with SQUID. The technique enhanced the sensitivity of measuring minimal residual disease over standard pathology methods for patients undergoing chemotherapy. http://www.aacr.org/ cli_gb.indd 1 7/04/09 18:30:53 10/8/09 10:33 AM Page 1 В© 2009 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries. RevcoFrigFreez_CLI_1109:Thermo How far would you go to protect your samples? Researchers worldwide protect more than two billion samples inside Thermo Scientific cold storage equipment. With +4В°C refrigerators to -196В°C cryogenic freezers and Thermo Scientific Nalgene and Nunc consumables, youвЂ™re free to concentrate on your work without worrying about your valuable samples. Thermo Scientific Revco high-performance laboratory refrigerators and freezers are engineered for the laboratory, providing: вЂў SecurityвЂ“Our IntrLogicв„ў microprocessor-based control system manages setpoint and actual conditions, providing the highest level of security вЂў EfficiencyвЂ“Advanced defrost sensors minimize frost built-up, maintaining coil efficiency вЂў VersatilityвЂ“A variety of sizes from undercounter to upright models is available to meet your space requirements So relax вЂ“ and choose Thermo Scientific Revco laboratory refrigerators and freezers. Visit www.thermo.com/cold Moving science forward www.cli-online.com & search 24853 Thermo Scientific Revco refrigerators and freezers are available for general and specialized applications including pharmacy, blood banking and chromatography. Visit us at Medica 2009 Dusseldorf/Germany вЂ“ November 18-21 Hall 01 / Both E11 вЂ“ Issue NВ°5 вЂ“ October 2009 34 Medica Preview NGAL assay for use on clinical chemistry analysers Over the past few years NGAL has become established as a new biomarker for diagnosing acute kidney injury (AKI). Using NGAL to diagnose AKI enables a marked improvement in the management of affected patients. Currently there is no preferable technology for the early diagnosis of AKI. Existing methods of determining kidney damage, e.g. the commonly used measurement of serum creatinine, only indicate renal failure resulting from a prior kidney injury at a relatively late stage (24-72 hours) after the injury has occurred, making this worthless for early diagnosis. Moreover, numerous studies have shown that NGAL responds earlier than all other renal status markers and shows a proportionate response to injury. NGAL, therefore, permits the early diagnosis and prognostic stratification of acute kidney injury and is expected to enable new specific therapies to be developed. Until now only ELISA-based NGAL assays have been commercially available, and the lack of a more suitable assay platform has impeded the translation of this promising biomarker from research into routine diagnostics, but NGAL will soon be available for use on clinical chemistry analysers. BioPorto Diagnostics is expanding its NGAL portfolio with the forthcoming launch of a turbidimetic NGAL test that is suitable for use with both blood and urine samples. The test is designed for routine diagnostic use on a wide range of automated clinical chemistry analysers that are already running in central laboratories in hospitals all over the world. Like the majority of other high volume routine biomarkers, the new NGAL test is based on the principle of turbidimetry, recognised to combine high diagnostic performance with fast turnaround time and therefore implicitly addressing the acute aspect of NGAL evaluation. BioPorto Diagnostics A/S Gentofte, Denmark Medica stand Hall 03/K88 www.cli-online.com & search 24817 Slide scanner for optimal histological examinations With its unprecedented scanning speed and top-quality on-screen imaging, the new Leica SCN400 Slide Scanner offers an alternative to microscopy for the examination of histological samples in pathology, research and teaching. The custom tailored lens for a digital sensor, specially designed for high-res scans, ensures that the resolution and colour fidelity of the image on the screen are just as good as that of the microscope image. Thanks to the dynamic focus principle which keeps the sample in focus for the full duration of the scan, even difficult samples can be effortlessly digitised. The slide scanner is able to load and scan up to four specimens at a time. With a scanning rate of 100 secs per 15 x 15 mm at 20x magnification, sample throughput is substantially increased. The matching Autoloader Leica SL801 is capable of scanning up to 384 samples at the same time, overnight if required, offering completely new options for automated operation. The user can keep loading new samples or remove finished scans without interrupting the process. Once a sample has been digitised, it can be easily retrieved, processed and made available to a defined group of users. Leica Microsystems GmbH Wezlar, Germany Medica Stand Hall 10/C32 www.cli-online.com & search 24818 Light resistant tubes Light-sensitive substances such as certain vitamins and enzymes, metallic salts, photosensitisers and antibodies coupled with fluorescent dyes must be stored under special conditions in the laboratory to ensure they are not degraded by UV radiation, leading to a loss of functionality. Existing brown glass flasks and polypropylene tubes can effectively protect these solutions and reagents from the harmful effects of light. However, they are not generally suitable for storing smaller sample sizes in the millilitre range. The portfolio of Light Protection Tubes for storing light-sensitive materials and light-sensitive reactions, which includes light-protection centrifuge tubes in 15 and 50 mL sizes, has been augmented by a 1.5 mL light-protection polypropylene reaction tube with attached cap. This is a major improvement for scientists who, until now, had to wrap smaller vessels in aluminium foil. Compared to transparent standard tubes, products made from pigmented polypropylene display no light transmission in a broad wavelength range to ensure optimal protection of the light-sensitive sample. Greiner Bio-One GmbH Frickenhausen, Germany Medica stand Hall 03/B70 www.cli-online.com & search 24736 www.cli-online.com & search 24810 Medica Preview Fully automated clinical chemistry analysers The Analyticon Biolyzer 200 is a fully automated clinical chemistry benchtop analyser with a throughput of up to 450 tests/hour. The system requires minimal space, while offering a broad range of parameters and easy-tooperate software, and is therefore a convenient solution for small to medium sized laboratories. A second clinical chemistry analyser will be launched in the first quarter of 2010 with an even higher throughput. The Biolyzer 600 will extend the companyвЂ™s line of analysers, covering medium sized laboratories or even the large laboratory as a specialised chemistry system and backup solution. A broad panel of parameters is available for both analysers as well as an optional direct ISE-unit for measuring electrolytes. Both systems use the same reagent concept, taking advantage of nearly 30 years of the companyвЂ™s experience in producing high-quality clinical chemistry reagents. ANALYTICON BIOTECHNOLOGIES AG Lichtenfels, Germany 35 вЂ“ Issue NВ°5 вЂ“ October 2009 Analyser for blood samples The CellaVision DM1200 automates the work that is traditionally carried out by laboratory personnel using microscopes. The analyser performs a preliminary WBC differential and RBC characterisation, and provides functionality for platelet estimation. Using technology for digital image analysis, cells in blood can be classified automatically, reducing the time taken and providing more standardised analyses. Final verification can be performed by medical technologists on their computer screens. The results are then sent automatically to the LIS for archiving. The digital analysis allows results to be shared and discussed with colleagues within or outside the hospital. The analyser is compatible with all other products from the company, and can easily become part of any haemotology workflow. CellaVision AB Lund, Sweden Medica Stand Hall 01/A32 www.cli-online.com & search 24802 Medica Stand Hall 3/E67 www.cli-online.com & search 24842 ISE module for clinical chemistry analyser Designed using automated 2-point calibration for measuring sodium, potassium and chloride in whole blood, serum and diluted urine, the ISE module can be customised for the userвЂ™s clinical chemistry analyser easily and flexibly. Results are available within 30 seconds, and the newly developed high sensitive electrodes and module allow calibration to be carried out only once a day. Very accurate results are obtained with a C.V of less than 0.5%. Jokoh Co Ltd Tokyo, Japan Medica Stand Hall 01/B26 www.cli-online.com & search 24800 Filter tips for safer pipetting and greater accuracy The unique SafetySpace filter tips have more space between the sample and the filter than conventional filter tips usually have, thus the user does not need to worry about the sample touching the filter regardless of the pipetting technique or type of liquid being handled. Accurate and precise results are obtained even when pipetting foaming liquids such as buffers and proteins. Unlike with other filter tips, these tips are also suitable for reverse pipetting as well as for multiple dispensing with electronic pipettes. Designed to meet high quality and purity demands, these tips are ideal for molecular biology, microbiology and cell culture applications, as well as radioactive work. The tip range covers seven different sizes from 10 ВµL up to 1200 ВµL. Packed in colour-coded single tray boxes, and certified DNase, RNase and endotoxin free as well as pre-sterilised, the tips are compatible with all Biohit pipettes and most other pipette brands. Biohit Ltd Helsinki, Finland Medica Stand Hall 02/A49 www.cli-online.com & search 24812 www.cli-online.com & search 24792 Medica Preview Cystatin C assay The range of tests available on MODULAR Analytics and cobas platforms for the assessment of glomerular filtration rate (GFR) has been extended with the launch of a new assay for the early assessment of glomerular filtration rate (GFR) in patients with impaired renal function. The Roche Cystatin C assay is more sensitive than creatinine measurements, allowing diagnosis of chronic kidney disease (CKD) at an early stage when therapeutic intervention is possible. Unlike creatinine, cystatin C levels are not affected by muscle mass, age (below 50), gender or inflammation and cystatin C is only eliminated via filtration, making it a convenient and accurate marker for the assessment of GFR. By indicating a reduction in GFR sooner, it is ideal for the assessment of patients in the early stages of CKD and reduces the need to perform invasive determinations of GFR. The Roche Cystatin C assay is promoted for use in the assessment of patients under the age of 60, for early diagnosis of initially minor kidney damage (for example in diabetes patients). It can also be used in monitoring kidney disease and post-transplantation patients, and for dose adjustment of renally excreted drugs. The assay is easy to use, requires minimal handling, and produces highly precise and sensitive results from low sample volumes (2ВµL). Roche Diagnostics GMBH Penzberg, Germany 37 it is easy to disassemble for cleaning and servicing. Five models are available, with adjustable volumes ranging from 0.25mL to 60mL. High precision and accuracy is ensured through several stages of strict quality checks during the manufacturing process. Each instrument is individually calibrated in accordance with ISO 8655 standards and comes with an individual calibration certificate. Microlit Bottle top dispensor Responding to current needs, the Microlit bottle top dispensor is a unique combination of affordable pricing and high performance. The simple design together with a variety of useful features facilitates all routine dispensing needs. The plunger movement is smooth and effortless. Each instrument comes with four adapters to fit most laboratory reagent bottles. The safety discharge system reduces the risk of accidental dispensing and the nozzle cap prevents any unwanted drops landing on the work space. The instrument is fully autoclavable at 121 В°C, 15psi, and whilst there is no need for disassembling the instrument for autoclaving, Medica Preview Designer antigen-based ELISAs A new milestone in the serological diagnosis of coeliac disease DNA sequence coding for Gliadin analogous peptides Medica Stand Hall 03/K36 www.cli-online.com & search 24843 Upright microscope with built-in LED illumination Offering consistent longterm performance for biological and medical applications, the costeffective CX21LED clinical microscope with builtin LED illumination can illuminate samples with a similar light intensity to that provided by halogen bulbs. Furthermore, the LED light source produces a more uniform, controllable and stable illumination for high-quality imaging. As an environmentally-friendly and easy-to-use system, this microscope is also ideal for educational use. OLYMPUS LIFE SCIENCE EUROPA GMBH Hamburg, Germany Medica Stand Hall 10/C20, 15/G12 www.cli-online.com & search 24809 Rapid test for TB The FASTSURE TB DNA rapid test is an accurate, simple, speedy and crosscontamination proof nucleic acid detection kit for the qualitative detection of Mycobacterium tuberculosis (TB) DNA in human samples, and a clinical diagnostic tool for TB infection. The test kit includes sample preparation, nucleic acid isothermal amplification & hybridisation, and finally detection using an innovative, cross-contaminationproof, user-friendly device that easily fits into the human hand. Superior sensitivity of has been demonstrated compared to solid media culture and smear microscopy. MP Biomedicals Santa Ana, CA, USA Medica Stand Hall 03/H56 www.cli-online.com & search 24745 Peptide 1 Peptide 2 GAF(gliadin analogous fusion peptide) GAF(3X) Lucknow, India Medica Stand Hall 02/A07 www.cli-online.com & search 24742 вЂ“ Issue NВ°5 вЂ“ October 2009 Chemical synthesis Sequence optimisation and assembly GAF-1 GAF-2 Expression in E. coli GAF-3 The Anti-Gliadin (GAF-3X) ELISA is based on a state-of-the-art recombinant gliadin-analogue fusion peptide, and provides significantly higher sensitivity and specificity than conventional anti-gliadin ELISAs. At 95% specificity the new ELISA showed a sensitivity of 83%/94% (IgA/ IgG), compared to 54%/31% (IgA/IgG) for a conventional ELISA. The Anti-Gliadin (GAF3X) ELISA (IgG) is especially suited for detection of patients with IgA deficiency, which is frequently associated with coeliac disease. A new standard in the diagnosis of WegenerвЂ™s granulomatosis The Anti-PR3-hn-hr ELISA employs an optimised antigen combination of human native (hn) and a designer, human expressed recombinant (hr) proteinase 3, which provides an unsurpassable antigen spectrum and unrivalled sensitivity for the detection of anti-PR3 autoantibodies. In a clinical study, 94% of cANCA reactive sera (indirect immunofluorescence) showed a positive reaction in the AntiPR3-hn-hr ELISA, compared with 88% for capture ELISA and 78% for conventional ELISA. A novel ELISA to aid diagnosis of primary biliary cirrhosis TP LB BCOADH-E2 PDH-E2 TP LB PBC spec. LB PBC spec. 138 aa 118 aa (His)g OGDH-E2 TP LB PBC spec. 82 aa BPO(3E) The Anti-M2-3E ELISA is based on a designer fusion protein, which contains all three relevant enzyme complexes (3E) of the mitochondrial antigen M2 (AMA M2): pyruvate dehydrogenase, branched-chain 2-oxoacid dehydrogenase and oxoglutarate dehydrogenase. In a clinical study the Anti-M2-3E ELISA showed a previously unattained sensitivity of 93%, compared to 79% for a conventional anti-M2 ELISA based solely on pyruvate dehydrogenase. The specificity of the new test system was 98%. EUROIMMUN AG Luebeck, Germany Medica Stand Hall 1, Booth A08 www.cli-online.com & search 24835 вЂ“ Issue NВ°5 вЂ“ October 2009 Medica Preview 38 Custom-made horseradish peroxidase and glucose oxidase BBI Enzymes has launched a new custom specification service which offers horseradish peroxidase and glucose oxidase tailor-made exclusively for customers. The efficient service creates the enzymes specifically to each customerвЂ™s own unique requirements, and promises batch to batch consistency, optimised isoenzyme content, and high purity and specificity. This service is the first launch from the companyвЂ™s new state of the art facility following a significant investment into BBI Enzymes. Promising a fast turnaround and guaranteeing highly competitive pricing, the service comes backed by BBI EnzymesвЂ™ 50 years of expertise and is ideal for anyone looking for a guaranteed enzyme supply. BBI Enzymes Blaenavon, Gwent, UK susceptibility to HIV. Many physicians do not routinely complete an examination unless a possible pregnancy is involved and yet this is the most common form of vaginal disease. The standard Gram stain test is, however, time consuming. A 60 second cost effective diagnostic tool for a quick, qualified, quality diagnosis of bacterial vaginosis, the VGTest, is now available. This new diagnostic tool will enable specialists, general practitioners and laboratories dedicated to womenвЂ™s healthcare, to quickly test, diagnose and so provide appropriate on the spot treatment. Based on ion mobility spectrometry technology, already a proven technology for detecting drugs, chemicals and explosives, the VGTest shows excellent laboratory results and provides a correct diagnosis within 60 seconds. The test detects elevated levels of trimethylamine (TMA), which is present in cases of infection or dysfunction. 3QBD Arad, Israel Medica Stand Hall 03/E44 www.cli-online.com & search 24845 Medica Stand Hall 1/F24 www.cli-online.com & search 24797 Rapid diagnosis of bacterial vaginosis Between 10-64% of the female population worldwide is at risk of the vaginal condition bacterial vaginosis at any given time. The disease very often remains asymptomatic and is thus undetected. BV can cause gynaecological and obstetric complications, including miscarriages, preterm births, pelvic inflammatory disease (PID), chorioamnionitis, puerperal endometritis and increased Anti-human adiponectin Mabs Adiponectin is an important regulator of lipid and glucose metabolism with anti-diabetic, antiinflammatory and anti-atherogenic properties. The potential diagnostic use of adiponectin has been a subject of increasing interest in recent years. There are several oligomeric forms of native adiponectin circulating in the blood, namely low-, medium- and high-molecular weight forms. Biological activity of adiponectin is mediated by the high-molecular weight form (HMW) and, not surprisingly, it has been suggested recently that concentration of the HMW form of adiponectin or the ratio HMW/total adiponectin (sum of three types of oligomers) in serum correlates more strongly with insulin resistance and other measures of type 2 diabetes than total adiponectin. A new generation of anti-human adiponectin monoclonal antibodies suitable both for research purposes (Western blotting, direct ELISA) and for the development of adiponectin-specific sandwich immunoassays are now available, as well as native antigen purified from human plasma. Hytest ltd Turku, Finland Medica Stand Hall 03/F44 www.cli-online.com & search 24738 Expanded range of quality controls Acusera, RandoxвЂ™ range of quality control sera, has expanded to include more new controls. The controls now include: blood gas; coagulation; drugs of abuse; liquid cardiac; liquid clinical chemistry; maternal screening; tumour markers and urinalysis. These assayed quality controls are highly accurate, and are supplied with instrument and method-specific values. Bottles, caps and packaging are all colour-coded to help distinguish different analyte levels and minimise costly errors. The company also offers customised quality control materials tailored to the customerвЂ™s specifications allowing uncompromised performance. Randox Laboratories Crumlin Co. Antrim, UK Mecica Stand Hall 03/A08 www.cli-online.com & search 24743 Software for analysis of flow cytometer data A new version of the software for the analysis of data acquired with FlowCytomix, a technology for multiple analyte detection on a flow cytometer, is available. FlowCytomix Pro 2.3 is very user-friendly, and incorporates many new features. Analysis is even more precise, since the new software offers the option to assay single samples and standards, duplicates or triplicates. File information is retained for further analysis, and the loading of files is faster. Individual settings can be saved at different stages, facilitating subsequent analyses. All plots generated in the course of an analysis can be saved and printed for improved data preparation. Bender MedSystems GmbH Vienna, Austria Medica Stand Hall 03/F56 www.cli-online.com & search 24793 www.cli-online.com & search 24741 Anuncio CLI 23/2/09 11:43 PпїЅgina 1 C M Y CM MY CY CMY K Just rest and we will do the rest Save on Problems... ... Choose Quality CLINICAL CHEMISTRY AUTOMATED SYSTEMS AUTOIMMUNITY QUALITY CONTROL RAPID TESTS INFECTIOUS IMMUNOLOGY Clean room for autoimmunity tests, Biosystems Biosafety laboratory for infectious immunology tests, BioSystems Costa Brava 30, 08030 Barcelona (Spain) Tel. +34-93 311 00 00 Fax +34-93 346 77 99 [email protected] вЂў www.biosystems-sa.com www.cli-online.com & search 24529 вЂ“ Issue NВ°5 вЂ“ October 2009 Product News 40 Single and multi channel pipettes Fully autoclavable and UV resistant, the Smart range of Accumax pipettes includes both single and multi channel models. An over-moulded TPE grip, sensitive 4 digits counter, Teflon sealing and unique locking system facilitate use. The ergonomic design and engineering ensure light weight handling, smooth plunger action and ease in operation. This range of pipettes is subject to the same stringent quality management system which made Accumax the first and only micropipette manufacturer in Asia-Pacific to have a calibration laboratory accredited with the highest standard for calibration: ISO 17025. www.cli-online.com & search 24795 Accumax Gandhinagar, Gujarat, India Medica Stand Hall 3/K66 www.cli-online.com & search 24846 Hepcidin ELISA A test kit is now available that facilitates the timely diagnosis of iron disorders, including haemochromatosis and anaemia; this diagnostic kit measures the level of the hormone hepcidin. In recent years, scientists have discovered that hepcidin helps regulate the amount of iron in humans. An unbalanced iron level can lead to many common medical conditions including anaemia and iron overload diseases, or it can occur in chronic kidney disease, inflammation, or diabetes mellitus. The DRG Hepcidin 25 (C-Terminal) ELISA Kit will be available to medical professionals worldwide, providing the first simple, fast and accurate method to test patient hepcidin levels. The results offer more information to clinicians to facilitate the diagnosis and treatment of medical conditions including iron deficiency diseases, some of the most common diseases worldwide. The kit provides accurate information and is user-friendly. DRG International Mountainside, NJ, USA www.cli-online.com & search 24794 www.cli-online.com & search 24811 Lab systems for mid-volume clinical laboratories The Dimension Vista 500 Intelligent Lab System is the latest addition to SiemensвЂ™ family of ultra-integrated chemistry and immunochemistry systems. The new system has a test menu of more than 115 assays and offers test panels for anaemia, cardiac disease, thyroid disorders, therapeutic drug monitoring, protein testing, drugs-of-abuse testing and routine and specialty chemistry testingвЂ”all on one system and from a single patient sample. With the option of two configurations, the system meets the varying needs of mid-volume laboratories: throughput is up to 1,000 patient tests per hour. The Dimension Vista 1000T Intelligent Lab System is also available by seamlessly adjoining two Dimension Vista 500 Systems, doubling the throughput capacity to up to 2,000 tests per hour and providing mirror back-up capabilities and increased walk-away time for laboratory personnel. Identical to the other Dimension Vista systems, four advanced detection technologies including photometry, nephelometry, V-LYTE electrolyte and LOCI advanced chemiluminescence are used on the systems, resulting in a broad test menu. The LOCI technology allows access to fast, sensitive immunoassay testing and a fast analytical turnaround time of 10 minutes for critical cardiac tests such as high-sensitivity Troponin I. Siemens Healthcare Diagnostics Newark, DE, USA www.cli-online.com & search 24748 Buprenorphine Enzyme Immunoassay Beckman Coulter has expanded its drugs of abuse test menu to include an enzyme immunoassay for the semi-synthetic opioid, buprenorphine. The test provides fast screening for the qualitative and semi-quantitative determination of norbuprenorphine (buprenorphine metabolite) in human urine. Reagents and calibrators for the assay are liquid and ready-to-use, eliminating the need Product News to mix, hydrate or pre-dilute reagents before testing. The assayвЂ™s cutoff value is 10ng/mL for norbuprenorphine, and it delivers analytical sensitivity of 3ng/mL for buprenorphine and norbuprenorphine (the active, dealkylated metabolite of buprenorphine). It cross-reacts at 94% with buprenorphine. Like all the companyвЂ™s Drugs of Abuse Test assays, this test is formulated to deliver the critical elements of effective DAT analysis, namely speed, accuracy, ease-of-use and economy. When used in conjunction with Synchron UniCel Clinical Chemistry Systems and i class Integrated Systems, this high quality assay provides timely, reliable and efficient DAT results. Beckman Coulter, Inc Nyon, Switerland www.cli-online.com & search 24844 HA LT for automated measurement of hyaluronic acid 41 вЂ“ Issue NВ°5 вЂ“ October 2009 Semi-automatic analyser Using state-ofthe-art technology, the BTS-350 new generation semi-automatic analyser is based on LED technology. It embraces concepts such as low energy consumption, low toxicity and a practically unlimited lifetime. Currently this is the only semi-automatic analyser that functions on a complete range of LEDs, has a USB drive to allow archiving and data transfer and a battery pack that not only functions as a back up when electricity fails, but also allows the analyser to be used anywhere at anytime. Furthermore, with hard coated filters, advanced ergonomics and improved optics, as well as low energy operation, the analyser requires basically no maintenance. In addition to the new and improved hardware technology, an intuitive, straightforward and user-friendly software encompasses all of the parameters necessary for routine assays with an excellent quality control scheme. An ideal solution to low-to-midsize laboratories worldwide, this analyser is flexible, reliable and optimised for IVD and laboratory testing. BioSystems SA Barcelona, Spain www.cli-online.com & search 24814 DiatronвЂ™s Reagent Division offers high quality reagents for Sysmex, Mindray, Beckman Coulter, Abbott, ABX, Erma hematology analyzers вЂў Long stability вЂў Proven reliability вЂў Cyanide free вЂў 20 years of experience вЂў High quality standards: A unique new test for the automated measurement of Hyaluronic Acid (HA) in serum and plasma using standard biochemical analysers is now available. Patient samples are mixed with a recombinant HA binding protein (HABP) in the HA LT reagent. Latex particles coated by anti-HABP antibody are added resulting in increasing turbidity. The high molecular weight glycosaminoglycan HA is the best single biomarker for hepatic fibrosis. It shows a negative predictive value (NPV) of up to 100% for cirrhosis of various aetiologies. Several biomarker combinations have been validated to enable the accurate staging of different fibrosis stages as non-invasive alternatives to liver biopsies. Fibrometer and Hepascore are the most powerful scores that include HA in their panels. HA LT by Wako can significantly improve such scores in terms of automation, simplicity and analytical performance. The test is highly precise, accurate and offers a high linear range from 10 to 1000 ng/mL. Whole genome amplification kit Current single-cell WGA kits using molecular displacement amplification (MDA) technology do not offer reproducible amplification of whole genomes, causing sporadic allele and locus dropouts, which seriously compromise results in Preimplantation Genetic Diagnosis (PGD). Hence, microarray and qPCR genetic analysis of single cells using MDAamplified DNA is noisy and produces unreliable data. The PicoPlex Single Cell Whole Genome Amplification (WGA) kit allows researchers to get the same amount of genomic information from one cell as they do from 10,000 cells. This technology enables labs to begin qPCR, microarray or sequencing analysis less than three hours after sample collection. The kit reproducibly amplifies total DNA one million-fold from single cells to produce five micrograms of amplified DNA in less than three hours. Wako Chemicals GmbH Rubicon Genomics Inc. Neuss, Germany Ann Arbour, MI, USA www.cli-online.com & search 24815 www.cli-online.com & search 24807 ISO 9001 and ISO 13485 вЂў Produced in EU вЂў Reasonable price For more information please contact us at [email protected] www.diatron.com Visit us on Medica 2009 Hall 3, Stand A07 www.cli-online.com & search 24641 BOOK REVIEWs 42 вЂ“ Issue NВ°5 вЂ“ October 2009 Surgical Pathology of Endocrine and Neuroendocrine Tumors Ed. by Ashraf Khan, Pub. by Springer (Humana), 2009, 242pp, в‚¬ 149 Surgical pathology is the cornerstone in the management of neoplastic disorders. Written for the practicing surgical pathologist in mind, this book, one of the Current Clinical Pathology series, provides an up-todate text on surgical pathology of endocrine and neuroendocrine tumours. The text begins with radiological imaging of tumours, followed by a section on fine needle aspiration biopsy. The main section focuses on surgical pathology of endocrine and neuroendocrine tumours. The volume closes with applications of molecular techniques and their potential for the future. Written by a panel of internationally recognised pathologists who are experts in their fields, the authors contribute their own valuable insights into making this new book an essential resource for practicing general and specialist surgical pathologists and pathologists in training. Leukemia Methods and Protocols Ed. by Chi Wai Eric So Pub. by Springer (Humana), 2009, 428pp, в‚¬74,95 The molecular basis and pathogenesis of leukaemia have been clarified, but better strategies are needed to diagnose, classify and treat this biologically and clinically diverse disease. Here experts bring together a wide range of state-of-the-art laboratory methods and detailed protocols that are useful for both clinical and basic research scientists working on the disease. The volume provides techniques for: prenatal backtracking of leukaemic clones; molecular diagnosis; detection of genome-wide genetic abnormalities and profiling; identification of unknown fusion genes; monitoring of minimal residual diseases; disease modelling using murine and human primary haematopoietic cells; studying of normal and malignant haematopoiesis; identification of interacting partners with leukaemia-associated oncoproteins; and global characterisation of genomewide epigenetic changes in leukaemic cells. Clinical and Diagnostic Virology by Goura Kudesia and Tim Wreghitt Pub. by Cambridge University Press 2009, 276pp, ВЈ29.99 This basic but comprehensive text is aimed at all healthcare professionals who need a clear understanding of medical virology. Written by two highly experienced virologists, the book is divided into five sections, namely individual viruses; other related agents; clinical syndromes; diagnostic techniques; and patient management. The individual virus chapters provide information on incubation period, infectivity, control of infection and management. The clinical syndrome chapters provide information on the clinical presentation of disease, thus enabling the reader to search according to patient symptoms rather than referring to several individual virus chapters. The standard chapter formats, simple language and liberal use of tables, figures and algorithms enable quick access to key information, and the comprehensive coverage of all viral agents is unique in a practical guide of this size. Springer Springer Cambridge University Press New York, NY, USA New York, NY, USA Cambridge, UK www.cli-online.com & search 24847 www.cli-online.com & search 24687 www.cli-online.com & search 24848 Calendar of events November 4-6, 2009 JournГ©es Internationales de Biologie (JIB) Paris, France Syndicat des Biologistes & Reed Expositions France Tel. +33 1 47 56 50 79 Fax +33 1 47 56 52 58 e-mail : [email protected] www.jib-sdbio.fr HEMATOLOGY REAGENTS FOR CELLCOUNTERS LABEX is a very experienced producer of reagents for various types of blood cell counters. All products are CE-marked and marketed world-wide through exclusive distributors. For further information about products, your local supplier or distribution partnership, please contact LABEX. Welcome to our new stand location Hall 1/F29 At MEDICA 2009 DГњSSELDORF th st 18 -21 NOV. 2009 P.O. Box 22159 SE-250 23 Helsingborg SWEDEN Tel +46 42 32 40 00 Fax +46 42 20 27 71 www.labex.com [email protected] www.cli-online.com &&search www.cli-online.com search00000 24767 November 5-7, 2009 7th Annual World Congress on Insulin Resistance San Francisco, CA, USA Tel. +1 818 342 1889 Fax +1 818 342 1538 e-mail: insulinresistance@ pacbell.net www.insulinresistance.us November 18-20, 2009 ESCMID Conference on Enterococci: from Animals to Man Barcelona, Spain European Society of Clinical Microbiology and Infectious Diseases Tel. +41 61 686 7799 Fax +41 61 686 7798 e-mail: [email protected] www.escmid.org November 18-21, 2009 MEDICA 2009 DГјsseldorf, Germany e-mail: [email protected] www.medica.de November 19-22, 2009 AMP 2009 Kissimmee, FL, USA Association for Molecular Pathology Tel. +1 301 634 7939 Fax +1 301 6347990 e-mail: [email protected] www.amp.org/2009/ December 11-13, 2009 MedifestвЂ™09 Pragati Maidan, New Delhi, India Vantage Trade Fairs (P) Limited Tel. +91 11 3058 0444 / 3058 0777 Fax +91 11 3058 1000 e-mail: [email protected] www.vantagemedifest.com/ medifest_india January 16-17, 2010 Melanoma 2010: 20th Annual Cutaneous Malignancy Update San Diego, CA, USA Scripps Health Tel. +1 858-652-5400 Fax +1 858-652-5565 e-mail: [email protected] www.scripps.org/events/ melanoma-annual-cutaneousmalignancy-update January 25-28, 2010 MEDLAB at Arab Health 2010 Dubai, United Arab Emirates IIR Middle East Tel. +971 4 3365161 Fax +971 4 3364021 www.arabhealthonline.com/ NEW_MEDLAB.html Microbiology & Infectious Disease Vienna, Austria Congrex Switzerland Ltd Tel. +41 61 686 77 11 Fax +41 61 686 77 88 e-mail: [email protected] www.congrex.ch/eccmid2010/ February 25-28, 2010 Early Disease Detection and Prevention (EDDP) conference 2010 Munich, Germany Paragon Conventions Tel. +41 22 5330 948 Fax +41 22 5802 953 e-mail: [email protected] www.paragon-conventions. com/eddp2010/ April 18-21, 2010 63rd CMEF Spring 2010 Shenzen, China Reed Sinopharm Exhibitions Tel. +86 10 6202 8899 ext 3825 Fax +86 20 6235 9314 e-mail: [email protected] http://en.cmef.com.cn/ February 27- March 2, 2010 The Role of Telomeres and Telomerase in Cancer research Fort Worth, TX, USA American Association for Cancer Research Tel. +1 215 440 9300 Fax +1 215 440 9313 e-mail: [email protected] www.aacr.org/page17420. aspx April 10-13, 2010 ECCMID 2010 вЂ“ 20th European Congress of Clinical May 9-13, 2010 Focus 2010 Glasgow, UK Association for Clinical Biochemistry www.focus-acb.org.uk For more events see: www.cli-online.com/events/ Dates and descriptions of future events have been obtained from official industrial sources. CLi cannot be held responsible for errors, changes or cancellations. Are we going in the right direction? Is my lab going in the right direction? The new IMMULITE 2000 XPi helps ensure that your lab is on the right path with its broad menu and connectivity options. With more than 20 years of proven immunoassay performance, the new IMMULITEВ® 2000 XPi* System maximizes productivity and reliability. As with all IMMULITEВ® systems, it features a wide range of routine, allergy and specialty assays, while easily connecting to automation and informatics platforms. Find out which IMMULITE system will place your lab on the right path at www.siemens.com/immunoassay Answers for life. A91DX-9054-A1-4A00 www.cli-online.com & search 24782 * Under development. Not available for sale in the U.S. В© 2009 Siemens Healthcare Diagnostics Inc. All rights reserved. IMMULITE is a trademark of Siemens Healthcare Diagnostics Inc. Working Together for Better Diagnostics Enzyme Immunoassay Kits from Fujirebio Diagnostics, Inc. Fujirebio Diagnostics is a world leader in in vitro diagnostics and the gold standard manufacturer of cancer biomarker assays worldwide. Features and Benefits of Fujirebio EIA Kits H E4 E I A M E SOMAR KВ® E IA Simple and robust protocols Ready to use reagents Simple one and two-step sandwich assays Ca n Ag Pr o G R P E I A C a n Ag S 1 0 0 E I A Streptavidin coated mircroplates Proprietary, characterized, high affinity monoclonal antibodies Ca n Ag CA 2 4 2 в„ў E I A Low sample volume C a n Ag S C C E I A See us at Medica Stand 03 F41 В© 2008 Fujirebio Diagnostics, Inc. EIA Kit, April 2008 www.cli-online.com & search 24784 www.FDI.com