SYNCHRON System(s)
Chemistry Information Sheet
PAM
Pancreatic Amylase
© 2015 Beckman Coulter, Inc. All rights reserved.
969650
For In Vitro Diagnostic Use
Rx Only
ANNUAL REVIEW
Reviewed by
Date
Reviewed by
Date
PRINCIPLE
INTENDED USE
PAM reagent, when used in conjunction with UniCel® DxC 600/800 System(s), is intended for the quantitative
determination of pancreas-specic amylase activity in human serum, plasma, or urine.
CLINICAL SIGNIFICANCE
Measurement of α-amylase is useful in the diagnosis and treatment of pancreatitis. The elevation of α-amylase activity,
however, is not specically indicative of pancreatic disorder, since the enzyme is also produced by the salivary gland and
other organs. Clinical evaluations of patients with acute pancreatitis have shown that pancreatic amylase has a greater
sensitivity than total amylase.1
METHODOLOGY
PAM reagent is used to measure pancreas-specic amylase by an immuno-inhibition method.
In the rst
incubation step, the activity of human salivary α-amylase is inhibited by two monoclonal antibodies which do not
affect pancreatic α-amylase. After a second incubation with the substrate, the α-amylase cleaves the substrate
(4,6-Ethylidene-G7p-Nitrophenol) into fragments and these fragments are further hydrolyzed by α-glucosidase to yield
p-nitrophenol and glucose.2
The SYNCHRON System(s) automatically proportions the appropriate sample and reagent volumes into the cuvette. The
ratio used is one part sample to 24 parts reagent. The system monitors the change in absorbance at 410 nanometers.
This change in absorbance is directly proportional to the activity of amylase in the sample and is used by the System to
calculate and express the pancreatic PAM activity.
Chemistry Information Sheet A18535 AN
DECEMBER 2015
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PAM
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CHEMICAL REACTION SCHEME
5 moles of substrate yields 5 moles of PNP
ET = Ethylidene
PNP = p-Nitrophenol
G = Glucose
SPECIMEN
TYPE OF SPECIMEN
Biological uid samples should be collected in the same manner routinely used for any laboratory test.3 Freshly drawn
serum or plasma, or freshly collected urine are the preferred specimens. Acceptable anticoagulants are listed in the
PROCEDURAL NOTES section of this chemistry information sheet. Whole blood is not recommended for use as a
sample.
SPECIMEN STORAGE AND STABILITY
1.
Tubes of blood are to be kept closed at all times and in a vertical position. It is recommended that the serum or
plasma be physically separated from contact with cells within two hours from the time of collection.4
2.
Separated serum or plasma should not remain at room temperature longer than 8 hours. If assays are not
completed within 8 hours, serum or plasma should be stored at +2°C to +8°C. If assays are not completed within
48 hours, or the separated sample is to be stored beyond 48 hours, samples should be frozen at -15°C to -20°C.
Frozen samples should be thawed only once. Analyte deterioration may occur in samples that are repeatedly
frozen and thawed.4
3.
It is recommended that urine assays be performed within 2 hours of collection.5 Pancreatic amylase in urine is
stable for ten days at +2°C to +8°C or two days at +20°C to +25°C. Pancreatic amylase is very unstable in acidic
urine. Specimens should be pH adjusted to approximately pH 7 prior to refrigeration.6 For timed specimens, the
collection container should be kept in the refrigerator or on ice during the timed period. Collect specimen without
additives. If the sample is turbid, or contains particulate matter, clarify by centrifugation (3000 x g for 10 minutes).
Additional specimen storage and stability conditions as designated by this laboratory:
SAMPLE VOLUME
The optimum volume, when using a 0.5 mL sample cup, is 0.3 mL of sample. For optimum primary sample tube volumes
and minimum volumes, refer to the Primary Tube Sample Template for your system.
PAM
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CRITERIA FOR UNACCEPTABLE SPECIMENS
Refer to the PROCEDURAL NOTES section of this chemistry information sheet for information on unacceptable
specimens.
Criteria for sample rejection as designated by this laboratory:
PATIENT PREPARATION
Special instructions for patient preparation as designated by this laboratory:
SPECIMEN HANDLING
Special instructions for specimen handling as designated by this laboratory:
REAGENTS
CONTENTS
Each kit contains the following items:
Two PAM Reagent Cartridges (2 x 60 tests)
VOLUMES PER TEST
Sample Volume
10 µL
ORDAC Sample Volume
3 µL
Total Reagent Volume
240 µL
Cartridge Volumes
A
– –
B
200 µL
C
40 µL
Chemistry Information Sheet A18535 AN
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PAM
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REACTIVE INGREDIENTS
REAGENT CONSTITUENTS
Reagent Mixture:
14.2 mL
α-Glucosidase (yeast, ≥4 kU/L)
Monoclonal antibodies (Murine source, 97 mg/L)
Starter Reagent:
4,6-Ethylidene-G7p-Nitrophenol (22 mmol/L)
3.3 mL
Also non-reactive chemicals necessary for optimal system performance.
NOTICE
Avoid all contact with reagent. Sweat and saliva contain α-amylase. It is recommended
that gloves be worn when handling the reagent cartridges. Use caution when
recapping reagent cartridges. Reagent Mixture and Starter Reagent caps must not be
interchanged or reagent contamination will occur.
GHS HAZARD CLASSIFICATION
Not classied as hazardous
Safety Data Sheet is available at techdocs.beckmancoulter.com.
MATERIALS NEEDED BUT NOT SUPPLIED WITH REAGENT KIT
At least two levels of control material
Saline
REAGENT PREPARATION
No preparation is required.
ACCEPTABLE REAGENT PERFORMANCE
The acceptability of a reagent is determined by ensuring that quality control results are within your facility’s acceptance
criteria.
REAGENT STORAGE AND STABILITY
PAM reagent, when stored unopened at +2°C to +8°C, will remain stable until the expiration date printed on the cartridge
label. Once opened, the reagent cartridge is stable for 30 days at +2°C to +8°C unless the expiration date is exceeded.
DO NOT FREEZE.
Reagent storage location:
PAM
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Chemistry Information Sheet A18535 AN
DECEMBER 2015
CALIBRATION
CALIBRATOR REQUIRED
Calibration is not required.
TRACEABILITY
This measurand (analyte) is traceable to the manufacturer’s selected Measurement Procedure as described in the
Methodology section.
QUALITY CONTROL
At least two levels of control material should be analyzed daily. In addition, these controls should be run with each new
reagent cartridge and after specic maintenance or troubleshooting procedures as detailed in the appropriate system
manual. More frequent use of controls or the use of additional controls is left to the discretion of the user based on good
laboratory practices or laboratory accreditation requirements and applicable laws. Controls used should contain both
active salivary amylase and pancreatic amylase to ensure the integrity of the reagent.
The SYNCHRON System(s) pancreatic amylase (PAM) reagent is designed to test for human pancreatic amylase.
Control materials consisting of porcine amylase do not perfectly mimic the performance of the reagent with patient
samples. Due to human pancreatic amylase specicity, noticeable lot to lot shifts with animal based control materials
may occur. To aid you in ensuring consistent reagent quality for human samples you may run several known patient
samples on both the new and old lots. Alternatively, you may contact the Clinical Support Center at 1-800-854-3633 from
the United States and Canada or your local Beckman Coulter Representative for the results of human patient samples
performed during manufacture.
The following controls should be prepared and used in accordance with the package inserts. Discrepant quality control
results should be evaluated by your facility.
Table 1.0 Quality Control Material
CONTROL NAME
SAMPLE TYPE
STORAGE
TESTING PROCEDURE(S)
1.
If necessary, load the reagent onto the system.
2.
Program samples and controls for analysis.
3.
After loading samples and controls onto the system, follow the protocols for system operations.
For detailed testing procedures, refer to the SYNCHRON LX Operations Manual, or the UniCel DxC 600/800 System
Instructions For Use (IFU) manual.
Chemistry Information Sheet A18535 AN
DECEMBER 2015
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PAM
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CALCULATIONS
The SYNCHRON System(s) performs all calculations internally to produce the nal reported result. The system will
calculate the nal result for sample dilutions made by the operator when the dilution factor is entered into the system
during sample programming.
REPORTING RESULTS
Equivalency between the SYNCHRON LX and UniCel DxC 600/800 Systems has been established. Chemistry results
between these systems are in agreement and data from representative systems may be shown.
REFERENCE INTERVALS
Each laboratory should establish its own reference intervals based upon its patient population. The reference intervals
listed below were taken from literature.7,8,9,10
Table 2.0 Reference intervals
INTERVALS
Literature
SAMPLE TYPE
CONVENTIONAL UNITS
S.I. UNITS
Serum or Plasma (+37°C)
≤ 46 U/L
≤ 0.8 µkat/L
Urine (+37°C)
< 320 U/L
< 5.3 µkat/L
SAMPLE TYPE
CONVENTIONAL UNITS
S.I. UNITS
INTERVALS
Laboratory
Refer to References (11,12,13) for guidelines on establishing laboratory reference intervals.
Additional reporting information as designated by this laboratory:
PROCEDURAL NOTES
ANTICOAGULANT TEST RESULTS
The following anticoagulants were assessed by Deming regression analysis with a minimum of 50 paired serum and
plasma samples. Values of serum (X) ranging from 13 U/L to 51 U/L were compared with the values for plasma (Y)
yielding the following results:
Table 3.0 Acceptable Anticoagulant Test Results
ANTICOAGULANT
LEVEL OF ANTICOAGULANT TESTED
DEMING REGRESSION ANALYSIS
Lithium Heparin
14 Units/mL
Y = 0.999X - 0.7; r = 0.993
Sodium Heparin
14 Units/mL
Y = 0.986X - 0.3; r = 0.982
1.5 mg/mL
Y = 0.954X - 0.4; r = 0.994
EDTA
PAM
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LIMITATIONS
1.
The recovery of pancreatic α-amylase activity is 99-100%. The residual activity of salivary α-amylase is
approximately 3%. In rare cases, elevated pancreatic α-amylase values may be obtained as a result of extremely
high activities of salivary α-amylase (>1000 U/L).
2.
Serum pancreatic amylase results on patients with macroamylase may be elevated. The elevation is not due to
the inability of the anti-salivary antibodies to inhibit the salivary amylase found in the serum immune complex, but
from the higher than normal level of pancreatic amylase which is in the immune complex. This elevated serum
pancreatic amylase is not diagnostic for pancreatitis.14,15,16
3.
Do not perform this test on grossly hemolyzed samples.
INTERFERENCES
1.
The following substances were tested for interference with this methodology:
Table 4.0 Interferences
SUBSTANCE
SOURCE
LEVEL TESTED
OBSERVED EFFECT
RBC hemolysate
500 mg/dL
NSIa
Bilirubin
Porcine
30 mg/dL
NSI
Lipemia
Human
500 mg/dL (4+)
NSI
Hemoglobin
a
NSI = No Signicant Interference (within ±10 U/L or 7%).
2.
Refer to References (17,18,19) for other interferences caused by drugs, disease and preanalytical variables.
3.
For assays employing mouse antibodies, the possibility exists for interference by human anti-mouse antibodies
(HAMA) in the sample. Human anti-mouse antibodies may be present in samples from patients who have received
immunotherapy or diagnostic procedures utilizing monoclonal antibodies or in individuals who have been regularly
exposed to animals.20,21 Additionally, other heterophile antibodies, such as human anti-goat antibodies may be
present in patient samples. Interpretation of results should be done in the context of the overall clinical presentation
of the patient, including symptoms, clinical history, data from additional tests and other appropriate information.
PERFORMANCE CHARACTERISTICS
ANALYTIC RANGE
NOTICE
Due to the use of different amylase substrate in this reagent as compared to the LX
Amylase Reagent, there is no direct conversion of units between total amylase and
pancreatic amylase.
The SYNCHRON System(s) method for the determination of this analyte provides the following analytical ranges:
Table 5.0 Analytical Range
SAMPLE TYPE
Serum/Plasma/Urine
Serum/Plasma/Urine (ORDAC)a
a
CONVENTIONAL UNITS
S.I. UNITS
7 – 600 U/L
0.12 – 10 µkat/L
480 – 1800 U/L
8 – 30 µkat/L
Overrange Detection and Correction. Refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual for more details on this function.
Samples with activities exceeding the high end of the analytical range should be rerun with ORDAC enabled or diluted
with saline and reanalyzed.
Chemistry Information Sheet A18535 AN
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REPORTABLE RANGE (AS DETERMINED ON SITE):
Table 6.0 Reportable Range
SAMPLE TYPE
CONVENTIONAL UNITS
S.I. UNITS
SENSITIVITY
Sensitivity is dened as the lowest measurable concentration which can be distinguished from zero with 95% condence.
Sensitivity for PAM determination is ≤7.0 U/L.
EQUIVALENCY
Equivalency was assessed by Deming regression analysis of patient samples to accepted clinical methods.
Serum or plasma (in the range of 10 to 548 U/L):
Y (SYNCHRON LX Systems)
= 1.026X + 3.4
N
= 164
MEAN (SYNCHRON LX Systems)
= 99.0
MEAN (SYNCHRON CX Systems PAMY)
= 93.2
CORRELATION COEFFICIENT (r)
= 1.000
Urine (in the range of 9 to 573 U/L):
Y (SYNCHRON LX Systems)
= 1.021X + 0.9
N
= 122
MEAN (SYNCHRON LX Systems)
= 196.3
MEAN (SYNCHRON CX Systems PAMY)
= 191.4
CORRELATION COEFFICIENT (r)
= 1.000
Refer to References (22) for guidelines on performing equivalency testing.
PRECISION
A properly operating SYNCHRON System(s) should exhibit precision values less than or equal to the following:
Table 7.0 Precision Values
TYPE OF
PRECISION
Within-run
PAM
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CHANGEOVER VALUEa
1 SD
SAMPLE TYPE
U/L
µkat/L
U/L
µkat/L
% CV
Serum/Plasma/Urine
5.0
0.08
143
2.29
3.5
Serum/Plasma/Urine
(ORDAC)
b
NA
NA
NA
10.0
NA
EN
Chemistry Information Sheet A18535 AN
DECEMBER 2015
Table 7.0 Precision Values, Continued
Total
a
b
CHANGEOVER VALUEa
1 SD
TYPE OF
PRECISION
SAMPLE TYPE
U/L
µkat/L
U/L
µkat/L
% CV
Serum/Plasma/Urine
7.5
0.12
143
2.29
5.3
Serum/Plasma/Urine
(ORDAC)
NA
NA
NA
NA
15.0
When the mean of the test precision data is less than or equal to the changeover value, compare the test SD to the SD guideline given above to
determine the acceptability of the precision testing. When the mean of the test precision data is greater than the changeover value, compare the
test % CV to the guideline given above to determine acceptability. Changeover value = (SD guideline/CV guideline) x 100.
NA = Not applicable.
Comparative performance data for the SYNCHRON LX® System evaluated using the NCCLS Approved Guideline EP5-A
appears in the table below.23 Each laboratory should characterize their own instrument performance for comparison
purposes.
Table 8.0 NCCLS EP5-A Precision Estimate Method
TYPE OF
IMPRECISION
Within-run
Total
a
SAMPLE TYPE
No.
Systems
No. Data
Pointsa
Test Mean
Value (U/L)
EP5-A Calculated
Point Estimates
SD
%CV
Serum
Control 1
1
80
64
0.8
1.3
Serum
Control 2
1
80
410
3.3
0.8
Serum
(ORDAC)
Control 3
1
80
772
8.3
1.1
Urine
Control
1
80
148
1.0
0.7
Serum
Control 1
1
80
64
2.7
4.3
Serum
Control 2
1
80
410
4.5
1.1
Serum
(ORDAC)
Control 3
1
80
772
10.5
1.4
Urine
Control
1
80
148
1.9
1.3
The point estimate is based on the pooled data from one system, run for twenty days, two runs per day, two observations per run on an instrument
operated and maintained according to the manufacturer’s instructions.
NOTICE
These degrees of precision and equivalency were obtained in typical testing procedures
on a SYNCHRON LX® System and are not intended to represent the performance
specications for this reagent.
ADDITIONAL INFORMATION
For more detailed information on UniCel DxC Systems, refer to the appropriate system manual.
Beckman Coulter, the Beckman Coulter Logo, Synchron, UniCel and DxC are trademarks of Beckman Coulter, Inc and
are registered in the USPTO.
SHIPPING DAMAGE
If damaged product is received, notify your Beckman Coulter Clinical Support Center.
Chemistry Information Sheet A18535 AN
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REVISION HISTORY
Revision AG
Revised Quality Control section.
Revision AH
Updated corporate address.
Revision AJ
Added Revision History.
Revision AK
Added new language requirement: Czech, and Korean.
Revision AL
Removed references to CX and LX systems as they are discontinued effective 12/2013.
Added Beckman Coulter trademark statement and disclaimer.
Revision AM
Added GHS Classication information
Revision AN
Added new language requirement: Romanian
FOOTNOTES
Beckman Coulter‘s SYNCHRON ® and DECISION® Control Products contain salivary amylase.
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REFERENCES
1.
Tietz, N. W., et al., "Multicenter Evaluation of a Specific Pancreatic Isoamylase Assay Based on Double
Monoclonal-Antibody Technique", Clin. Chem., 34:2096 2102 (1998).
2.
Gerber, M., Wulff, K., Laboratoriums Medizin., 12:110 113 (1988).
3.
Tietz, N. W., "Specimen Collection and Processing; Sources of Biological Variation", Textbook of Clinical
Chemistry, 5th Edition, W. B. Saunders, Philadelphia, PA (2005).
4.
National Committee for Clinical Laboratory Standards, Procedures for the Handling and Processing of Blood
Specimens Approved Guideline, NCCLS publication H18-A, Villanova, PA (1990).
5.
National Committee for Clinical Laboratory Standards, Urinalysis and Collection, Transportation and Preservation
of Urine Specimens, Approved Guideline, Vol. 15, No. 15 , NCCLS publication GP16-A, Villanova, PA (1995).
6.
Tietz, N. W., Clinical Guide to Laboratory Tests, 3rd Edition, W. B. Saunders Company, Philadelphia, PA (1995).
7.
Junge, W., et al., Clinical Biochemistry, 22:109 114 (1989).
8.
Tietz, N. W., et al., Clin. Chem., 34:2096 2102 (1988).
9.
Hohenwallner, W., et al., J. Clin. Chem. Biochem., 27:97 101 (1989).
10.
Kruse-Jarres, J. D., et al., J. Clin. Chem. Biochem., 27:103 113 (1989).
11.
National Committee for Clinical Laboratory Standards, How to Define, Determine, and Utilize Reference Intervals
in the Clinical Laboratory, Approved Guideline, 2nd Edition, NCCLS publication C28-A, Villanova, PA (2000).
12.
Tietz, N. W., ed., Fundamentals of Clinical Chemistry, 6th Edition, W. B. Saunders, Philadelphia, PA (2007).
13.
Henry, J. B., Clinical Diagnosis and Management by Laboratory Methods, 18th Edition, W. B. Saunders,
Philadelphia, PA (1987).
14.
Tietz, N. W., Clinical Guide to Laboratory Tests, 3rd Edition, W. B. Saunders Company, pp 416 417, Philadelphia,
PA (1995).
15.
Klonoff, D., West J. Med, 133:392 407, Nov. (1980).
16.
Lott, J., "Inflammatory Diseases of the Pancreas", CRC Critical Reviews in Clinical Laboratory Sciences, 17/3:201
228.
17.
Young, D. S., Effects of Drugs on Clinical Laboratory Tests, 5th Edition, AACC Press, Washington, D. C. (2000).
18.
Friedman, R. B., Young, D. S., Effects of Disease on Clinical Laboratory Tests, 4th Edition, AACC Press,
Washington, D.C. (2001).
19.
Young, D. S., Effects of Preanalytical Variables on Clinical Laboratory Tests, 3rd Edition, AACC Press,
Washington, D. C. (2007).
20.
Bjerner, J., et al., "Immunometric Assay Interference: Incidence and Prevention", Clin. Chem. 48:613 621 (2002).
21.
Kricka, L. J., "Interferences in Immunoassays-Still a Threat", Clin. Chem., 46:1037 1038 (2000).
22.
National Committee for Clinical Laboratory Standards, Method Comparison and Bias Estimation Using Patient
Samples Approved Guideline, NCCLS publication EP9-A, Villanova, PA (1995).
Chemistry Information Sheet A18535 AN
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23.
National Committee for Clinical Laboratory Standards, Precision Performance of Clinical Chemistry Devices, 2nd
Edition, Approved Guideline, Vol. 19, No. 2, NCCLS publication EP5-A, Villanova, PA (1999).
Beckman Coulter Eurocenter S.A., 22, rue Juste-Olivier. Case Postale 1044, CH - 1260 Nyon 1, Switzerland
Tel: +41 (0)22 365 36 11
Beckman Coulter, Inc., 250 S. Kraemer Blvd., Brea, CA 92821 U.S.A.
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