en

ALT
07P98
Alanine Aminotransferase Reagent Kit G71193R01
B7P980
Created October 2016.

07P9820

Instructions must be carefully followed. Reliability of assay results Safety Precautions

cannot be guaranteed if there are any deviations from these CAUTION: This product requires the handling of human specimens.
instructions. It is recommended that all human-sourced materials be considered
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NAME potentially infectious and handled in accordance with the OSHA
Alinity c Alanine Aminotransferase Reagent Kit (also referred to as Standard on Bloodborne Pathogens. Biosafety Level 2 or other
ALT) appropriate biosafety practices should be used for materials that
contain or are suspected of containing infectious agents.1-4
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INTENDED USE Safety Data Sheets are available at [Link] or
The Alinity c Alanine Aminotransferase assay is used for the contact your local representative.
quantitation of alanine aminotransferase in human serum or plasma For a detailed discussion of safety precautions during system
on the Alinity c analyzer. operation, refer to the Alinity ci-series Operations Manual, Section 8.
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SUMMARY AND EXPLANATION OF THE TEST Reagent Handling
Alanine Aminotransferase (ALT), also referred to as glutamate • Upon receipt, place reagent cartridges in an upright position
pyruvate transaminase (GPT), is an enzyme involved in amino for 1 hour before use to allow bubbles that may have formed to
acid metabolism. It is found in many tissues, but the highest levels dissipate.
are found in liver and kidney tissues. Tissue destruction leads to • If a reagent cartridge is dropped, place in an upright position
the release of the intracellular enzyme into the circulating blood. for 1 hour before use to allow bubbles that may have formed to
Markedly elevated serum ALT levels may be found in a variety of dissipate.
diseases which involve the liver, such as hepatitis, mononucleosis,
• Reagents are susceptible to the formation of foam and bubbles.
and cirrhosis. These very high levels of ALT are not usually observed
Bubbles may interfere with the detection of the reagent level in
in other disease processes, e.g., myocardial infarction; thus, ALT is
the cartridge and cause insufficient reagent aspiration that may
regarded as a reasonably specific indicator of liver disease.
adversely affect results.
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PRINCIPLES OF THE PROCEDURE For a detailed discussion of reagent handling precautions during
ALT present in the sample catalyzes the transfer of the amino group system operation, refer to the Alinity ci-series Operations Manual,
from L-alanine to α-ketoglutarate, forming pyruvate and L-glutamate. Section 7.
Pyruvate in the presence of NADH and lactate dehydrogenase (LD) Reagent Storage
is reduced to L-lactate. In this reaction NADH is oxidized to NAD.
Storage Maximum Additional Storage
The reaction is monitored by measuring the rate of decrease in Temperature Storage Time Instructions
absorbance at 340 nm due to the oxidation of NADH to NAD.
Unopened 2-8°C Until Store in upright position.
Methodology: Enzymatic: NADH (without P-5'-P) expiration
For additional information on system and assay technology, refer to date
the Alinity ci-series Operations Manual, Section 3. Onboard System 27 days
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REAGENTS Temperature
Opened 2-8°C Until Store in upright position.
Kit Contents
expiration Do not reuse original
Alinity c Alanine Aminotransferase Reagent Kit 07P98
date reagent caps or
Volumes (mL) listed in the table below indicate the volume per
replacement caps due to
cartridge.
the risk of contamination
07P9820 and the potential to
Tests per cartridge 360 compromise reagent
Number of cartridges per kit 10 performance.
Tests per kit 3600 Reagents may be stored on or off the system. If removed from the
68.1 mL system, store reagents with new replacement caps in an upright
21.0 mL position at 2 to 8°C. For reagents stored off the system, it is
recommended that they be stored in their original trays or boxes to
Active ingredients: β-NADH (0.16 mg/mL), Lactate ensure they remain upright.
dehydrogenase (2.57 U/mL), L-Alanine (392 mmol/L). For information on unloading reagents, refer to the Alinity ci-series
Active ingredients: α-Ketoglutaric acid (77 mmol/L), L-Alanine Operations Manual, Section 5.
(1000 mmol/L). Indications of Reagent Deterioration
Deterioration of the reagents may be indicated when a calibration
Warnings and Precautions error occurs or a control value is out of the specified range.
• Associated test results are invalid, and samples must be retested.
• For In Vitro Diagnostic Use Assay recalibration may be necessary.
• For troubleshooting information, refer to the Alinity ci-series
Operations Manual, Section 10.

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INSTRUMENT PROCEDURE Specimen Storage
The Alinity c Alanine Aminotransferase assay file must be installed Numerous publications have defined storage conditions for ALT.6-15
on the Alinity c analyzer prior to performing the assay. Examples are shown below.
For detailed information on assay file installation and viewing and Maximum
editing assay parameters, refer to the Alinity ci-series Operations Specimen Storage
Manual, Section 2. Type Temperature Time Special Instructions
For information on printing assay parameters, refer to the Alinity ci- Serum/ 30°C 3 days10 Remove serum or
series Operations Manual, Section 5. Plasma plasma from the clot,
For a detailed description of system procedures, refer to the Alinity red blood cells, or
ci-series Operations Manual. separator gel.
7 days10
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SPECIMEN COLLECTION AND PREPARATION 2 to 8°C
FOR ANALYSIS -40°C 60 days15
Specimen Types
It is recommended that specimens be assayed on the day of
The specimen types listed below were verified for use with this
collection.16, 17
assay.
When samples were stored at -20°C for 8 days, an 11% reduction
Other specimen types, collection tube types, and anticoagulants
in ALT activity was observed; a 20% reduction in ALT activity was
have not been verified with this assay.
observed when specimens were stored at -20°C for 1 month.18
Specimen Types Collection Tubes Special Conditions
Avoid multiple freeze/thaw cycles.
Serum Serum tubes (with or Stored specimens must be inspected for particulates. If present, mix
without gel barriers) with a low speed vortex or by inversion and centrifuge the specimen
Plasma Collection tubes Do not use to remove particulates prior to testing.
Acceptable ammonium heparin.5
anticoagulants are:
Specimen Shipping
Package and label specimens in compliance with applicable state,
Lithium heparin (with
federal, and international regulations covering the transport of clinical
or without gel barrier)
specimens and infectious substances.
Sodium heparin
EDTA ll
PROCEDURE
Hemolysis in serum or plasma can increase test results. Materials Provided
CAUTION: Erythrocytes contain approximately 3 to 5 times more ALT 07P98 Alinity c Alanine Aminotransferase Reagent Kit
than does serum.6 Materials Required but not Provided
• The instrument does not provide the capability to verify specimen • Alinity c Alanine Aminotransferase assay file
types. It is the responsibility of the operator to verify that the • Commercially available controls containing alanine
correct specimen types are used in the assay. aminotransferase
Specimen Conditions • Saline (0.85% to 0.90% NaCl) for specimen dilution
• For accurate results, serum and plasma specimens should be For information on materials required for operation of the instrument,
free of fibrin, red blood cells, and other particulate matter. Serum refer to the Alinity ci-series Operations Manual, Section 1.
specimens from patients receiving anticoagulant or thrombolytic For information on materials required for maintenance procedures,
therapy may contain fibrin due to incomplete clot formation. refer to the Alinity ci-series Operations Manual, Section 9.
• For accurate results, plasma specimens should be free of Assay Procedure
platelets and other particulate matter. Ensure centrifugation is For a detailed description of how to run an assay, refer to the Alinity
adequate to remove platelets. ci-series Operations Manual, Section 5.
• To prevent cross contamination, use of disposable pipettes or • If using primary or aliquot tubes, refer to the Alinity ci-series
pipette tips is recommended. Operations Manual, Section 4 to ensure sufficient specimen is
Preparation for Analysis present.
• Follow the tube manufacturer’s processing instructions for • To minimize the effects of evaporation, verify adequate sample
collection tubes. Gravity separation is not sufficient for specimen cup volume is present prior to running the test.
preparation. • Minimum sample volume requirements:
• Specimens should be free of bubbles. Remove bubbles with an –– Sample volume for single test: 5.3 µL.
applicator stick before analysis. Use a new applicator stick for NOTE: This amount does not include the dead volume
each specimen to prevent cross-contamination. plus the additional over-aspiration volume. For total sample
To ensure consistency in results, recentrifuge specimens prior to volume requirements, refer to the Alinity ci-series Operations
testing if Manual, Section 4.
• they contain fibrin, red blood cells, or other particulate matter. • Refer to the commercially available control material package
NOTE: If fibrin, red blood cells, or other particulate matter are insert for preparation and usage.
observed, mix by low speed vortex or by inverting 10 times prior to • For general operating procedures, refer to the Alinity ci-series
recentrifugation. Operations Manual, Section 5.
• For optimal performance, it is important to perform routine
maintenance as described in the Alinity ci-series Operations
Manual, Section 9. Perform maintenance more frequently when
required by laboratory procedures.

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Sample Dilution Procedures ll
RESULTS
Samples with an alanine aminotransferase value exceeding 3899 U/L Calculation
are flagged with the code "> 3899 U/L" and may be diluted with
The Alinity c Alanine Aminotransferase assay utilizes the Factor data
either the Automated Dilution Protocol or the Manual Dilution
reduction method to generate a calibration curve and results. The
Procedure.
calibration factor for the Alinity c Alanine Aminotransferase assay is
Automated Dilution Protocol 8141.
The system performs a 1:5 dilution of the sample and automatically
calculates the concentration by multiplying the result by the dilution Flags
factor. Some results may contain information in the Flags field. For a
Manual Dilution Procedure description of the flags that may appear in this field, refer to the
Alinity ci-series Operations Manual, Section 5.
Dilute the sample with saline (0.85% to 0.90% NaCl).
The operator must enter the dilution factor in the Specimen or Measuring Interval
Control tab of the Create Order screen. The system will use this Measuring interval is defined as the range of values in U/L which
dilution factor to automatically calculate the concentration of the meets the limits of acceptable performance for linearity, imprecision,
sample and report the result. and bias.
If the operator does not enter the dilution factor, the result must be The measuring interval of the Alinity c Alanine Aminotransferase
manually multiplied by the appropriate dilution factor before reporting assay is 5 to 3899 U/L.
the result. If a diluted sample result is less than the lower value of
the measuring interval of 5 U/L, do not report the result. Rerun using
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LIMITATIONS OF THE PROCEDURE
Refer to the SPECIMEN COLLECTION AND PREPARATION FOR
an appropriate dilution.
ANALYSIS and SPECIFIC PERFORMANCE CHARACTERISTICS
For detailed information on ordering dilutions, refer to the Alinity ci- sections of this package insert.
series Operations Manual, Section 5.
Calibration ll
EXPECTED VALUES
It is recommended that each laboratory determine its own reference
For instructions on performing a calibration, refer to the Alinity ci-
range based upon its particular locale and population characteristics.
series Operations Manual, Section 5.
Reference Range
Calibration is stable for approximately 27 days (648 hours), but
is required with each change in reagent lot. Verify calibration with Serum/Plasma20, 21
at least 2 levels of controls according to the established quality Range (U/L)
control requirements for your laboratory. If control results fall outside Adult 0 to 55
acceptable ranges, recalibration may be necessary.
This assay may require recalibration after maintenance to critical ll
SPECIFIC PERFORMANCE CHARACTERISTICS
parts or subsystems or after service procedures have been Representative performance data are provided in this section.
performed. Results obtained in individual laboratories may vary.
Quality Control Procedures The Alinity c analyzer, and the ARCHITECT c System and AEROSET
System utilize the same reagents and sample/reagent ratios.
As appropriate, refer to your laboratory standard operating
procedure(s) and/or quality assurance plan for additional quality Unless otherwise specified, all studies were performed on the Alinity
control requirements and potential corrective actions. c analyzer.
• Two levels of controls (normal and abnormal) are to be run every Precision
24 hours. Within-Laboratory Precision
• If more frequent control monitoring is required, follow the A study was performed based on guidance from CLSI
established quality control procedures for your laboratory. EP05-A2. Testing was conducted using 1 lot of Alinity c Alanine
• If quality control results do not meet the acceptance criteria Aminotransferase Reagent Kit, water calibrator, and 1 lot of
defined by your laboratory, sample results may be suspect. commercially available controls, and 1 instrument. Three controls
Follow the established quality control procedures for your were assayed in a minimum of 2 replicates at 2 separate times per
laboratory. Recalibration may be necessary. For troubleshooting day on 20 different days.22
information, refer to the Alinity ci-series Operations Manual, Within-Run Within-Laboratory
Section 10. (Repeatability) (Total)a
• Review quality control results and acceptance criteria following a Sample n Mean (U/L) SD %CV SD %CV
change of reagent or calibrator lot. Control 120 30 0.5 1.8 0.9 2.8
Level 1
Commercial controls should be used according to the guidelines
and recommendations of the control manufacturer. Concentration Control 120 108 0.7 0.6 1.0 1.0
Level 2
ranges provided in the control package insert should be used only for
Control 120 230 0.7 0.3 1.4 0.6
guidance. Level 3
For any control material in use, the laboratory should ensure that the a Includes within-run, between-run, and between-day variability.
matrix of the control material is suitable for use in the assay per the
assay package insert.
Quality Control Guidance
Refer to “Basic QC Practices” by James O Westgard, Ph.D. for
guidance on laboratory quality control practices.19
Verification of Assay Claims
For protocols to verify package insert claims, refer to Verification of
Assay Claims in the Alinity ci-series Operations Manual.

3
Lower Limits of Measurement 8. Heins M, Heil W, Withold W. Storage of serum and whole blood
A study was performed based on guidance from CLSI EP17-A2 and samples? Effects of time and temperature on 22 serum analytes. Eur
was replicated on 3 reagent lots and 2 instruments over a minimum J Clin Chem Clin Biochem 1995;33:231–238.
9. Dale JC, Pruett SK. Phlebotomy—a minimalist approach. Mayo Clin
of 3 days on each instrument/reagent lot combination. The maximum Proc 1993;68(3):249-255.
observed Limit of Blank (LoB), Limit of Detection (LoD) and Limit of 10. Young D. Effects of Preanalytical Variables on Clinical Laboratory
Quantitation (LoQ) values are reported in the table.23 Tests, 2nd ed. Washington, DC: AACC Press; 1997;3–12.
U/L 11. Elfath D, Cooney J, McDaniel R, et al. Effect of frozen storage of
LoBa 1 serum on the level of 22 chemistry analytes [Poster 0102 presented
at AACC 43rd National Meeting in Washington, DC: July 30, 1991].
LoDb 2 Clin Chem 1991;37(6):931.
LoQc 5 12. Faulkner AM, Lukes-Hall AM, White GW. Evaluation of the Grenier
a plasma separator blood tube. Ann Clin Biochem 1990;27:386-387.
The LoB represents the 95th percentile from n ≥ 60 replicates of 13. Wilding P, Zilva JA, Wilde CE. Transport of specimens for clinical
zero-analyte samples. chemistry analysis. Ann Clin Biochem 1977;14:301–306.
b The LoD represents the lowest concentration at which the analyte 14. Schwartz MK. Interferences in diagnostic biochemical procedures.
can be detected with 95% probability based on n ≥ 60 replicates of Adv Clin Chem 1973;16:10.
low-analyte level samples. 15. Mosley JW, Goodwin RF. Stability of serum glutamic pyruvic
c The LoQ was determined from n ≥ 60 replicates of low-analyte transaminase activity on storage. Am J Clin Pathol 1965;44:591-595.
16. Williams K, Williams A, Kline L, et al. Stability of serum alanine
level samples and is defined as the lowest concentration at which a aminotransferase activity. Transfusion 1987;27(5):431–433.
maximum allowable precision of 20 %CV was met. 17. Cuccherini B, Nussbaum S, Seeff L, et al. Stability of aspartate
Linearity aminotransferase and alanine aminotransferase activities. J Lab Clin
Med 1983;102(3):370-376.
A study was performed based on guidance from CLSI EP06-A.24 18. Donnelly JG, Soldin SJ, Nealon DA, et al. Stability of twenty-five
This assay is linear across the measuring interval of 5 to 3899 U/L. analytes in human serum at 22°C, 4°C, and -20°C. Pediatr Pathol Lab
Interference Med 1995;15:869-874.
19. Westgard JO. Basic QC Practices. 3rd ed. Madison, WI: Westgard
This study was performed on the AEROSET System. Quality Corporation; 2010.
Potentially Interfering Endogenous Substances 20. Kaplan LA, Pesce AJ, editors. Clinical Chemistry Theory, Analysis,
Interference studies were conducted using NCCLS EP7-P.25 and Correlation, 2nd ed. St Louis, MO: CV Mosby; 1989:895–898.
Interference effects were assessed by Dose Response and Paired 21. Sherman KE, Dodd RY, et al. Alanine aminotransferase levels among
Difference methods, at the medical decision level of the analyte. volunteer blood donors: geographic variation and risk factors. J Infect
Dis 1982;145(3):383-386.
Potentially Interferent Level 22. Clinical and Laboratory Standards Institute (CLSI). Evaluation of
Interfering Target Recovery Precision Performance of Quantitative Measurement Methods;
Substance Default Units Alternate Units U/L (% of Target) Approved Guideline—Second Edition. CLSI Document EP05-A2.
Bilirubin 30 mg/dL 513 μmol/L 53.1 95.3 Wayne, PA: CLSI; 2004.
60 mg/dL 1026 μmol/L 53.1 88.1 23. Clinical and Laboratory Standards Institute (CLSI). Evaluation of
Hemoglobin 750 mg/dL 7.5 g/L 47.4 107.9 Detection Capability for Clinical Laboratory Measurement Procedures;
1000 mg/dL 10.0 g/L 47.4 111.0 Approved Guideline—Second Edition. CLSI Document EP17-A2.
Wayne, PA: CLSI; 2012.
Intralipid 550 mg/dL 5.5 g/L 50.6 97.2 24. Clinical and Laboratory Standards Institute (CLSI). Evaluation of
625 mg/dL 6.25 g/L 50.6 96.8 the Linearity of Quantitative Measurement Procedures: A Statistical
Approach; Approved Guideline. CLSI Document EP06-A. Wayne, PA:
Interferences from medications or endogenous substances may CLSI; 2003.
affect results.26 25. National Committee for Clinical Laboratory Standards (NCCLS).
Method Comparison Interference Testing in Clinical Chemistry; Proposed Guideline.
NCCLS Document EP7-P. Villanova, PA: NCCLS; 1986.
A study was performed based on guidance from CLSI EP9-A3 using 26. Young DS. Effects of Drugs on Clinical Laboratory Tests, 4th ed.
the Passing-Bablok regression method.27 Washington, DC: AACC Press; 1995:3-16–3-22.
Correlation Concentration 27. Clinical and Laboratory Standards Institute (CLSI). Measurement
Units n Coefficient Intercept Slope Range Procedure Comparison and Bias Estimation Using Patient Samples;
Alinity c Alanine Serum U/L 130 1.00 1.11 0.97 6-3727 Approved Guideline—Third Edition. CLSI Document EP09-A3. Wayne,
Aminotransferase PA: CLSI; 2013.
vs ARCHITECT Note for number formatting:
Alanine • A space is used as thousands separator (example: 10 000
Aminotransferase
specimens).
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BIBLIOGRAPHY • A period is used to separate the integer part from the fractional
part of a number written in decimal form (example: 3.12%).
1. US Department of Labor, Occupational Safety and Health
Administration, 29 CFR Part 1910.1030, Bloodborne pathogens.
2. US Department of Health and Human Services. Biosafety in
Microbiological and Biomedical Laboratories. 5th ed. Washington, DC:
US Government Printing Office; December 2009.
3. World Health Organization. Laboratory Biosafety Manual. 3rd ed.
Geneva: World Health Organization; 2004.
4. Clinical and Laboratory Standards Institute (CLSI). Protection
of Laboratory Workers From Occupationally Acquired Infections;
Approved Guideline—Fourth Edition. CLSI Document M29-A4. Wayne,
PA: CLSI; 2014.
5. Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical Chemistry,
2nd ed. Philadelphia, PA: WB Saunders; 1994:795–797.
6. Henry RJ, Cannon DC, Winkelman JW. Clinical Chemistry Principles
and Technics, 2nd ed. Hagerstown, MD: Harper and Row; 1974:888.
7. Ruby SG, Relber NE, Lonser RE. Preanalytical variation in alanine
aminotransferase. Clin Chem 1988:34(4):744–745.

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Key to Symbols
Consult instructions for use

Manufacturer

Sufficient for

Temperature limitation

Use by/Expiration date

In Vitro Diagnostic Medical

Device
Lot Number
Produced for Abbott by
Product of Canada
Reagent 1
Reagent 2
List Number
For use by or on the order of a
physician only (applicable to USA
classification only).
Serial number

Alinity, ARCHITECT, and AEROSET are trademarks of Abbott

Laboratories in various jurisdictions. All other trademarks are property
of their respective owners.

Abbott GmbH & Co. KG

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65205 Wiesbaden
Germany
+49-6122-580

Sekisui Diagnostics P.E.I. Inc.

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Charlottetown
Prince Edward Island
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Customer Service: Contact your local representative
or find country-specific contact information on
[Link]
Created October 2016.
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