Juin 2016 – Modèle 24
Cisbio Bioassays
Parc Marcel Boiteux – BP 84175 – 30200 Codolet / France - Tél. 33 (0)4.66.79.67.00
GASTRIN
GASK-PR
Trousse pour le dosage radioimmunologique
de la gastrine humaine dans le sérum ou le plasma
Radioimmunoassay kit for the quantitative determination of gastrin in
human serum or plasma.
Pour diagnostic In Vitro
For In Vitro diagnostic use
La trousse contient :
Traceur  37 kBq
Antisérum
Calibrateur 0
Calibrateurs 1 - 5
Réactif immunoprécipitant
Sachet plastique
Notice d’utilisation
Kit content :
Tracer  37 kBq
Antiserum
Calibrator 0
Calibrators 1 – 5
Immunoprecipitating reagent
Plastic bag
Instruction for use
1 x 10 mL
1 x 30 mL
1 x 5 mL
5 x 1 mL
1 x 100 mL
1
1
1 x 10 mL
1 x 30 mL
1 x 5 mL
5 x 1 mL
1 x 100 mL
1
1
Attention: Certains réactifs contiennent de l’azoture de sodium
Warning: Some reagents contain sodium azide
Immunoradiometrischer Test zur quantitativen Bestimmung
von Humangestrin in Serum oder Plasma
Kit per il dosaggio radioimmunologico della gastrina umana
nel siero o nel plasma
Zur In Vitro Diagnostik
Per uso diagnostico In Vitro
Inhalt des Kits :
Tracer  37 kBq
Antiserum
Kalibrator 0
Kalibratoren 1 – 5
Immunpräzipitierendes reagenz
Plastikbeutel
Gebrauchsinformation
Contenuto del kit :
Tracciante  37 kBq
Antisiero
Calibratore 0
Calibratori 1 – 5
Reagente immunoprecipitante
Sacchetto di plastica
Istruzioni per l’uso
1 x 10 mL
1 x 30 mL
1 x 5 mL
5 x 1 mL
1 x 100 mL
1
1
Achtung: Einige Reagenzien enthalten Natriumazid
Attenzione: Alcuni reagenti contengono sodio azide
1
1 x 10 mL
1 x 30 mL
1 x 5 mL
5 x 1 mL
1 x 100 mL
1
1
FRA
ENG
Explication des
symboles
Explanation of
symbols
Conforme aux
normes
européennes
Ab
RIP
European
conformity
DEU
Erläuterung
der Sumbole
ITA
ELL
POL
HUN
Spiegazione
dei simboli
Eπεξήγηση
των
συμβόλων
Wyjaśnienie
symboli
Jelmagyarázat
Σύμφωνο προς
τα ευρωπαϊκά
πρότυπα
Zgodne z
normami
europejskimi
Conformita
CE-Konformitätskennzeichnung
europea
T° limite de
stockage
Storage
temperature
limitation
Limitierung der
Lagertemperatur
Limiti per la
temperatura di
conservazione
Όριο
θερμοκρασίας
αποθήκευσης
Graniczna
temperatura
przechowywania
N° de lot
Batch code
Chargencode
codice lotto
Αριθμός
παρτίδας
Numer partii
Utiliser jusqu’au
Use by
utilizzare entro
Ημερομηνία
λήξης
Zużyć do
Consulter la
notice d’utilisation
Consult operating
instructions
Diagnostic In
Vitro
In Vitro Diagnostic
device
Fabriqué par
Manufactured by
Référence
Catalogue number
Verwendbar bis
Das Handbuch
zu Rate ziehen
consultare le
istruzioni per
l‘USO
InVitroDiagnostisch
e Anwendung
Dispositivo
Diagnostico In
Vitro
Prodotto da
Hergestellt von
N. catalogo
Katalog Nr.
Nombre de tubes
Number of
determinations
Traceur radioactif
Radioactive tracer
Antisérum
Antiserum
Calibrateur
Calibrator
Immunoprecipitating
Réactif
reagent
immunoprécipitant
Megfelel az
európai
szabványoknak
Tárolási
hőmérséklethatár
CES
Vysvětlení
symbolů
Evropská
shody
RUS
TUR
Объяснение
символов
Sembollerin
açıklaması
Европейское
соответствие
Avrupa'ya uyum
Mezní teplota
skladování
Ограничение
температуры
хранения
Gyártási szám
Č. šarže
номер партии
Felhasználható az
alábbi dátumig :
Použitelné do
Šifra serije
дата
истечения
срока действия
Son kullanım
tarihi
Upotrebiti do
Přečtěte si
návod k
použití
Учитывать
Руководство
по
эксплуатации
İşletim
talimatlarına
danışın
Pogledajte
uputstvo za
upotrebu
Diagnostika
in vitro
In Vitro
диагностическ
ое устройство
In Vitro
Tanılama cihazı
Uređaj za
dijagnostiku in
vitro
Üretici
Proizveo
Katalog
numarası
Kataloški broj
Diagnostyka In
Vitro
In vitro
diagnosztika
Κατασκευάζεται
από την
Wyprodukowane
przez
Gyártja:
Wzorzec
Referenciakészít
mény
Reference
номер по
каталогу
Vyrobil
Изготовитель
Vysvetlenie
symbolov
Európska zhoda
Parti kodu
∆ιαγνωστική
συσκευή In
Vitro
SVK
Evropska
usaglašenost
Ograničenje
temperature
za čuvanje
Olvassa el a
használati
utasítást
Αριθμός
καταλόγου
Objašnjenje
simbola
Depolama
sıcaklığı
sınırlaması
Patrz dołączona
ulotka
Συμβουλευτείτε
τις οδηγίες
χρήσης
SRP
Limity teploty
skladovania
Kód šarže
Použiteľné do
Prečítajte si
návod na použitie
In vitro
diagnostická
pomôcka
Výrobca
Katalógové číslo
Počet stanovení
Anzahl der
Bestimmungen
Numero di
determinazioni
Αριθμός
προσδιορισμών
Liczba probówek
A kémcsövek
száma
Počet
zkumavek
Количество
определений
Saptama sayısı
Broj
određivanja
Radioactiver
Tracer
Tracciante
radioattivo
Ραδιενεργός
ιχνηθέτης
Znacznik
radioaktywny
Nyomjelző izotóp
Tracer
пробирки с
покрытием
Radyoaktif
izleyici
Radioaktivni
indikator
Rádioaktívny
značkovač
Antisiero
αντιορος
Antysurowica
Antiserum
Antisérum
иммунная
сыворотка
Anti-serum
Antiserum
Antisérum
Calibratore
Βαθμονομητής
Kalibrator
Kalibrátor
Kalibrátor
калибратор
Kalibratör
Kalibrator
Kalibrátor
Reagente
immuno
∆ιάλυμα
κατακρήμνισης
odczynnik
Immuno
Immuno
Imunoprecipit
ační činidlo
иммунопрецип
итации реагент
Presipitasyon
solüsyonu
Precipitirajući
rastvor
Imunoprecipitačn
é činidlo
Antiserum
Kalibrator
Immunpräzipitier
endes reagenz
precipitante
precypitujacy
2
precipitating
reagens
GASK-PR
MISE A JOUR / UPDATING
Cisbio Bioassays - Juin 2016 - Modèle 24
FRA
Modifications par rapport à la version précédente :
7.1 enlever ± 1% (précision) /12.4 Enlever protection HAMA
ENG
Changes from the previous version:
7.1 remove ± 1% (precision) / 12.4 Remove “HAMA protection”
DEU
Änderungen gegenüber der Vorgängerversion:
7.1 zu entfernen ± 1% (Präzision) / 12.4 „HAMA-Schutz“ entfernen
ITA
Modifiche rispetto alla versione precedente:
7.1 rimuovere ± 1% (precisione) / 12.4 Eliminata “protezione HAMA”
ELL
Αλλαγές από την προηγούμενη έκδοση:
7.1 Αφαίρεση του «± 1% (ακρίβεια)» / 12.4 Αφαίρεση του «Προστασία HAMA»
POL
Zmiany w stosunku do poprzedniej wersji:.
7.1 usunąć z precyzją+- 1% / 12.4 usunięcie „Ochrona HAMA”
HUN
Változások az előző verzióhoz képest:
7.1 távolítsa pontosságú ± 1% / 12.4 a védelem megszüntetése "HАМА"
CES
Změny od předchozí verze:
7.1 odstranit ± 1 % (přesnost) / 12.4 Odstranit „Ochrana proti HAMA“
RUS
Изменения по сравнению с предыдущей версией:
7.1 удалить 7,1 ± 1% (точность) / 12.4 Удалить «защита HAMA»
TUR
Bir önceki sürüm üzerinde yapılan değişiklikler:
7.1 ± %1 (hassasiyet) ifadesi çıkarıldı / 12.4 “HAMA koruması” ifadesi çıkarıldı
SRP
Promene od prethodne verzije:
7.1 ukloniti ± 1% (preciznost) / 12.4 Ukloniti “zaštita HAMA”
SVK
Zmeny oproti predchádzajúcej verzii:
7.1 odstrániť ± 1% (presnosť)/12.4 Odstrániť „Ochrana proti HAMA“
3
GASK-PR
ENG
Cisbio Bioassays - June 2016 - Model 24
1. NAME AND INTENDED USE
GASK-PR is a radioimmunoassay kit for the direct quantitative determination of gastrin in human serum or plasma.
2. INTRODUCTION
Gastrin is a gastro-intestinal hormone secreted by the G cells of the gastric antral mucosa and by the duodenum. There are several
forms of gastrin in the blood: big big gastrin, big gastrin (G 34), little gastrin (G 17) and mini gastrin (G 14). The biological activity
common to all these forms is bound to the C-terminal tetrapeptide. Plasmatic gastrin has a half-life of approximately 8 minutes, and its
metabolism is mainly renal.
The main factors controling gastrin secretion are antra mechanical distension, alimentary contact, central stimulation via the
pneumogastric nerve, gastrointestinal hormones and medicines. Antral pH is the main regulator of gastrin secretion, as it is
suppressed when the pH descends below 3.
Gastrin stimulates not only the secretion of gastric acid, but that of intrinsic factor, of bicarbonate secretion by the pancreas, and the
release of insulin and calcitonin. Indirectly, it stimulates pepsin secretion. It also has a trophic effect on the digestive mucosa and
plays a role in digestive motricity.
The main application of gastrin assays lies in the early diagnosis of Zollinger-Ellison syndrome, a gastrin-secreting tumor usually of
pancreatic origin. Such cases give a clear-cut gastrin reaction to the secretin test.
High basal gastrin levels have been reported in cases of antral G cell hyperplasia, of atrophic gastritis, of Biermer’s anemia and of
vagotomy.
Correct interpretation of gastrinemia requires a knowledge of the patient’s basal and post-stimulation gastric acid secretion, and of his
or her renal function and age, as well as of the conditions under which samples were taken.
3. PRINCIPLE
The principle of the assay is based on competition between gastrin radiolabeled with iodine 125 and gastrin contained in the
standards or samples to be assayed for a given limited number of anti-gastrin antibody sites.
At the end of the incubation period, the amount of radiolabeled gastrin bound to the antibody is inversely proportional to the amount of
non-radiolabeled gastrin originally present in the assay.
The method proposed for the separation of the free and bound fractions uses an immunoprecipitant in which a second antibody has
been preprecipitated in excess.
4. REAGENTS
Each kit contains enough reagents for 100 tubes. The expiry date is marked on the external label.
REAGENTS
125I GASTRIN: lyophilized. Synthetic gastrin 1 (1-17),
buffer, bovine albumin, red dye.  37 kBq ( 1 µCi).
Reconstitute with 10 mL of distilled water.
ANTISERUM: lyophilized. Anti-gastrin rabbit antiserum,
buffer, normal rabbit serum, bovine albumin, preservative,
blue dye. Reconstitute with 30 mL of distilled water.
CALIBRATORS: lyophilized. Synthetic gastrin 1 (1-17),
buffer, bovine albumin, sodium azide. 20 – 60 - 150 - 500 1500 µU/mL*. Reconstitute with 1 ml of distilled water.
SYMBOLS
TRACER
Ab
CAL
0 CALIBRATOR: lyophilized. Buffer, bovine albumin,
sodium azide. Reconstitute with 5 mL of distilled water.
CAL
IMMUNOPRECIPITATING REAGENT: ready for use.
Insoluble complex of anti-rabbit IgG sheep serum and nonimmunized rabbit IgG, buffer, polyethylene glycol, sodium
azide.
RIP
QUANTITY
1 qs
10 mL
vial
1 qs
30 mL
vial
5 qs
1 mL
vials
1 qs
5 mL
vial
1
100 mL
vial
STORAGE
2-8°C until the expiry date.
Use immediately after reconstitution:
not use beyond.
2-8°C until the expiry date.
Use immediately after reconstitution:
not use beyond.
2-8°C until the expiry date.
Use immediately after reconstitution:
not use beyond.
2-8°C until the expiry date.
Use immediately after reconstitution:
not use beyond.
do
do
do
do
2-8°C until the expiry date.
(*) The values shown above are only target values: the true value of each calibrator is shown on its label. 1 µU MRC 68/439 = 1.05 pg
MRC 68/439.
5. PRECAUTIONS FOR USE
5.1. Safety measures
Raw materials of human origin contained in the reagents of this kit have been tested with licensed kits and found negative for the antiHIV 1, anti-HIV 2, anti-HCV antibodies and the HBs antigen. However as it is impossible to strictly guarantee that such products will
not transmit hepatitis, the HIV virus, or any other viral infection, all raw materials of human origin including the samples to be assayed
must be treated as potentially infectious.
Do not pipette by mouth. Do not smoke, eat or drink in areas in which specimens or kit reagents are handled. Wear disposable gloves
while handling kit reagents or specimens and wash hands thoroughly afterwards. Avoid splashing.
Decontaminate and dispose of specimens and all potentially contaminated materials as if they contained infectious agents. The
recommended method for doing this is autoclaving for a minimum of one hour at 121.5°C.
Sodium azide may react with lead or copper piping to form highly explosive metal azides. During waste disposal, flush the drains
thoroughly to prevent a build-up of these products.
5.2. Basic radioprotection rules
This radioactive product may only be received, purchased, stored or used by persons so authorized, and by laboratories covered by
such authorization. The solution should under no circumstances be administered to humans or to animals.
The purchase, storage, use or exchange of radioactive products are subject to the laws in force in the user's country.
Enforcement of the basic radioprotection rules will ensure adequate security.
4
GASK-PR
ENG
Cisbio Bioassays - June 2016 - Model 24
A summary of these is given below:
Radioactive products must be stored in their original containers in a suitable area. A record of the reception and storage of radioactive
products must be kept up to date. Handling of radioactive products should take place in a suitably-equipped area with restricted
access (controlled zone). Do not eat, drink, smoke or apply cosmetics in a controlled zone. Do not mouth-pipette radioactive solutions.
Avoid any direct contact with all radioactive products by using laboratory coats and protective gloves. Contaminated laboratory
equipment and glassware must be disposed of immediately after contamination to prevent cross-contamination of different isotopes.
Any contamination or radioactive substance loss should be dealt with in accordance with the established procedures. All radioactive
waste disposal must be carried out according to the regulations in force.
5.3. Handling precautions
Do not use kit components beyond their expiry date. Do not mix reagents from different batches. Avoid any microbic contamination of
reagents or of the water used for the reconstitution of reagents. Before each incubation, well mix the content of the tubes to ensure an
homogeneous medium. Respect the recommended speed and length of centrifuging time in order to ensure the adherence of the
immunoprecipitate to the bottom of the tubes. The use of trays enabling the simultaneous rturning over of several tubes is
recommended. Draining on an absorbent support is necessary to obtain reproducible results (better duplicates, reduced non specific
activity). Do not turn the tubes over twice. Respect the specified temperature conditions.
6. SPECIMEN COLLECTION AND PREPARATION
The assay is performed directly on serum or plasma without heparin. If the assay is to be performed within 24 hours of sample
collection, the samples may be stored at 2-8°C. Otherwise, it is better to divide them into aliquots and store deep-frozen (-20°C).
Dilutions
Should elevated gastrin levels be suspected, the 0 calibrator found in the kit is used for dilution.
It is recommended that disposable plastic tubes be used when carrying out dilution.
7. ASSAY PROCEDURE
7.1. Equipment required
Precision micropipettes or similar with disposable tips, permitting the dispensing of 100 µL, 300 µL and 1000 µL. Their calibration
must be checked regularly.
Distilled water. Disposable plastic tubes. Vortex-type mixer. Multitube centrifuge (1500 g minimum), refrigerated if possible. Invertable
tube racks.
Gamma scintillation counter calibrated for 125 iodine measurement.
7.2. Protocol
The reconstitution of the reagents and their dispensing into the tubes is carried out at room temperature (18-25°C).
The assay requires the following groups of tubes:
T group for the determination of total activity,
0 group for the 0 point of the curve and the determination of binding capacity,
Calibrator groups to establish the standard curve,
Sx groups for the samples to be assayed.
It is recommended that the assays be performed in triplicate for the T, 0 and calibrator groups and in duplicate for samples.
Respect the order in which reagents are to be added:
Dispense 100 µL of calibrators or samples into the corresponding groups of tubes.
Add 100 µL of radiolabeled gastrin to all the tubes.
Add 300 µL of antiserum to the standard and sample tubes.
Gently mix each tube using a Vortex-type mixer.
Incubate for 2 hours at 18-25°C.
Manually shake the content of the immunoprecipitating reagent vial to ensure its homogeneous suspension.
At room temperature, dispense 1 ml of this reagent into all the tubes (except those of the T group).
This step must be carried out as rapidly as possible.
Mix and allow to incubate for 15 minutes at 18-25°C.
Centrifuge all tubes (except those of the T group) at 1500 to 2000 g for 15 minutes, if possible at 2-8°C.
Remove the supernatant either by aspiration or by decantating. The recommended method is to invert the tubes over an "active" sink
or a container suitable for holding radioactive solutions. Shake the tubes and leave them inverted for 10 minutes on an absorbent
support.
Measure the radioactivity of each tube with a gamma scintillation counter.
8. QUALITY CONTROL
Good laboratory practices require that control samples be used in each series of assays to check the quality of the results obtained.
These samples must be treated in exactly the same way as the samples to be assayed, and it is recommended that the results be
analyzed with appropriate statistical methods.
9. RESULTS
For each group of tubes, compute the mean counts after subtracting the background.
Evaluate the system's liaison capacity.
CAL "0"
(Bo/T) %
= ------------ x 100
T
Calculate the binding percentages of the standards and the samples compared to the 0 calibrator.
CAL ou Sx
(B/Bo) %
= -------------- x 100
CAL "0"
Draw the standard curve plotting the B/Bo% of the calibrators as a function of their concentration.
It is recommended to use semi-log coordinates.
5
GASK-PR
ENG
Cisbio Bioassays - June 2016 - Model 24
Other methods may be used for drawing the curve, in particular by plotting the cpm versus concentration.
Read the samples’ values directly from the curve, correcting the read value for the dilution factor if necessary.
Typical calibrator curve (example only): this data must under no circumstances be substituted for results obtained in the laboratory.
T
Calibrator 0
Calibrator 1
Calibrator 2
Calibrator 3
Calibrator 4
Calibrator 5
Mean
cpm
14371
6481
5671
4938
3629
1711
745
B/Bo
x 100
100
87.5
76.2
56.0
26.4
11.5
2074
32.0
Sample
Concentration
µU/mL
0
20
60
150
500
1500
100
80
B/BO x 100
Tube groups
440
60
40
20
0
10
100
G AS K
μU /m l
1 000
10. PROCEDURAL LIMITATIONS
Samples which show turbidity, haemolysis, hyperlipemia or contain fibrin may give misleading results.
Do not extrapolate sample values beyond the last calibrator. Dilute the samples concerned and re-assay.
11. EXPECTED VALUES
Each laboratory must establish its own range of normal values. Values shown below are given as an example only.
Number of
cases
75
Extreme values
(µU/mL)
28 - 185
Mean
(µU/mL)
68.5
97% of measured values are between 28 and 115 µU/ml.
12. SPECIFIC CHARACTERISTICS OF THE ASSAY
12.1. Imprecision
This was evaluated using 2 samples with different concentrations assayed either 30 times in the same series or in duplicate in 10
different series.
Sample
Mean (µU/mL)
Within-run
Between-run
(CV %)
(CV %)
1
45.8
8.9
9.9
2
480
8.4
5.9
12.2. Recovery test
Known quantities of gastrin were added to human serum. The recovery percentages for gastrin in the samples ranged between 95
and 105%.
11.3. Detection limit
The detection limit is defined as being the smallest detectable concentration different from 0 with a probability of 95%. It has been
assessed as being 10 µU/mL.
12.4. Interference
No interference with bilirubin and triglycerides, measured up to respective concentrations of equal to 250 mg/L and 20 g/L, has been
observed.
ASSAY FLOW CHART
Calibrators,
Samples
µL
125I
Gastrin
µL
Antiserum
µL
T
-
100
-
0
100
100
300
Calibrators
100
100
300
Sx
100
100
300
Tubes
Mix.
Incubate
2 hours
at 1825°C.
Immunoprecipitating
reagent (mL)
1
1
1
6
Mix.
Incubate
15 minutes
at 18-25°C.
Centrifuge for
15 minutes.
Discard the
supernatant
Count
BIBLIOGRAPHY :
Debas HT, Runsteen D, Lundberg PA, et al. On the natural history of hypergastrinemia. Clin
Chem. 1987;10/3:222-5.
Lindstedt G, Runsteen D, Lundberg PA, et al. On the natural history of hypergastrinemia. Clin
Chem. 1985;31/7:1135-40.
Pricus MR, Cartez RP, Chen J, et al. On the biologically active structures of chollcystokinin,
little gastrin and enkephalin in the gastro intestinal system. Proc Natl Acad Sci USA.
1987;84/14:4821-5.
Schubert ML, Shamburek RD. Control of acid secretion. Gastroentoral Clin North Am.
1990;19/1:1-25.
Slingerland DW, Cardarelli JA, Burrows BA, Miller A. The utility of serum gastrin levels in
assessing the significence of low serum B12 levels. Arch Intern Med. 1984;144/6:1167-8.
7