BIOLABO
www.biolabo.fr ALT GPT (IFCC) Single vial
MANUFACTURER: Reagent for quantitative determination of Alanine amino transferase activity (ALT)
BIOLABO SAS, [EC 2.6.1.2] in human serum or plasma.
Les Hautes Rives
02160, Maizy, France
REF 80027 R1: 20 X 10 mL REF 80127 R1: 8 x 30 mL
REF 80227 R1: 10 x 125 mL REF 80327 R1: 6 x 200 mL
|
TECHNICAL SUPPORT AND ORDERS
Tel: (33) 03 23 25 15 50
IVD IN VITRO DIAGNOSTIC USE
Fax: (33) 03 23 256 256
CLINICAL SIGNIFICANCE (1) (2) SAFETY CAUTIONS
ALT is present in very high amounts in liver and kidney, and in smaller BIOLABO reagents are designated for professional, in vitro diagnostic
amounts in skeletal muscle and heart. Although serum levels of both use.
AST and ALT become elevated whenever diseases process affecting • Verify the integrity of the contents before use.
liver cells integrity, ALT is the more liver-specific enzyme. Serum • Use adequate protections (overall, gloves, glasses).
elevations of • Do not pipette by mouth.
ALT activity is rarely observed in conditions other than parenchymal • In case of contact with skin and eyes, thoroughly wash affected
liver disease (cirrhosis, carcinoma, hepatitis, obstructive jaundice or areas with plenty of water and seek medical advice.
liver stroke). Moreover its elevation persists longer than do those of • Reagents contain sodium azide (concentration < 0.1%) which may
AST activity. Measurement of both AST and ALT has some value in react with copper and lead plumbing. Flush with plenty of water
distinguishing hepatitis from other parenchymal lesions. when disposing.
• Material Safety Data Sheet is available upon request.
• Waste disposal: Respect legislation in force in the country.
PRINCIPLE (4) (5) (6)
All specimens should be handled as potentially infectious, in
Method developed by Wrobleski and LaDue, optimised by Henry and accordance with good laboratory practices using appropriate
Bergmeyer (following modified IFCC recommendations). Reaction
precautions. Respect legislation in force in the country.
scheme is as follows:
ALT
L- Alanine + 2-Oxoglutarate Pyruvate + L-Glutamate STABILITY AND STORAGE
Store away from light, well cap in the original vial at 2-8°C
LDH
Pyruvate + NADH + H+ L-Lactate + NAD+ • Unopened, reagents are stable until expiry date stated on the label
of the kit when stored and used as described in the insert.
The decrease in absorbance due to the conversion of NADH into • Once reconstituted, working reagent is stable for 60 days when free
NAD+, and proportional to ALT activity in the specimen, is measured at from contamination.
340 nm.
• Discard any reagent if cloudy or if absorbance measured at 340 nm
Absence of P5P allows a better stability of working reagent. is < 1.000.
• Don’t use working reagent after expiry date stated on the label of the
REAGENTS COMPOSITION Kit.
Vial R1 WORKING REAGENT SPECIMEN COLLECTION AND HANDLING (2) (7)
2-Oxoglutarate 15 mmol/L Unhemolysed serum. Do not use heparinised plasma.
L-Alanine 500 mmol/L ALT is stable in serum or plasma for:
LDH > 1600 Ul/L • 24 hours at room temperature.
NADH < 0.18 mmol/L • 7 days at 2-8°C.
Tris Buffer 100 mmol/L
pH at 30°C 7.50 + 0.1 INTERFERENCES (3) (6)
Preservative Hemoglobin: No interference up to 300 µmol/L Hb.
Before reconstitution: Xn, Harmful
Hemolysis: Positive interference due to ALT released from
R22-32: Harmful if swallowed. Contact with acid libaerates very toxic gas. erythrocytes.
S22-S281: Do not breath dust. After contact with skin, rinse immediately with plenty of Bilirubin: No interference up to 20 mg/dl (342 µmol/L).
water Turbidity: No interference up to 7.00 mmol/L triglycerides.
Once reconstituted: None LDH contained in reagent allows, during pre-incubation step, the
reduction of endogenous pyruvate which would positively interfere.
REAGENTS PREPARATION Elevated ALT level may involve NADH depletion during pre-incubation
stage, which may lead to under-estimated results. In case of lipemic or
REF 80027: Use a non-sharp instrument to remove aluminium cap. icteric specimens, increased absorbance may mask this phenomenon.
Add promptly to the contents of the vial the amount of demineralised It’s recommended to check these specimens diluted (1 + 4) in saline
water stated on the label. solution.
Mix gently and wait for complete dissolution before using reagent
For a more comprehensive review of factors affecting this assay refer
(approximately 2 minutes).
to the publication of Young D.S.
MATERIAL REQUIRED BUT NOT PROVIDED
1. Medical analysis laboratory equipment.
2. Normal and pathological control sera.
3. Demineralised water for reagent preparation.
Made in France Latest revision: www.biolabo.fr Revision: 18/12/2014
CALIBRATION MANUAL PROCEDURE
Results will depend on the accuracy of the instrument calibration, the Let stand reagents and specimens at room temperature.
time counting, the respect of reagent/specimen ratio and the
temperature control. Pipette into 1 cm path length thermostated cuvette:
• Use the theoretical calibration factor (§ CALCULATION) Reagent 1 mL
• Or REF 95015 BIOLABO Multicalibrator (calibration value
determined with validated statistical technics and metrologically Bring to 37°C (30°C) then add:
controlled instrument)
• or a multiparametric calibrator traceable to a reference method or Specimen 100 µL
material
Mix. Start a timer. Record initial absorbance after 1 minute at 340 nm.
Record the absorbance again every minutes during 3 minutes.
QUALITY CONTROL Calculate absorbance change per minute (∆Abs/min).
• BIOLABO EXATROL-N Level I REF 95010
BIOLABO EXATROL-P Level II REF 95011 Note: Specific procedures are available upon request for
• Other assayed control sera referring to the same method. automated instruments. Please contact BIOLABO technical
support.
• External quality control program.
It is recommended to control in the following cases:
• At least once a run. CALCULATION
• At least once within 24 hours. Calculate the result as follows:
• When changing vial of reagent.
• After maintenance operations on the instrument.
If control is out of range, apply following actions: With theoretical factor:
1. Repeat the test with the same control.
2. If control is still out of range, prepare a fresh control serum and ∆ Abs/min) x 1746
IU/L = (∆
repeat the test.
3. If control is still out of range, verify analysis parameters:
Wavelength, temperature, specimen/reagent ratio, time counting, IU/L
calibration factor. µKat/L =
4. If control is still out of range, use a new vial of reagent and 60
reassay With seric multicalibrator:
5. If control is still out of range, please contact BIOLABO technical
support or your local Agent.
(∆Abs/min) Assay
ALT Activity = x Calibrator Concentration
EXPECTED VALUES (2) (∆Abs/min) Calibrator
UI/L at 30°C at 37°C
Newborns, Infants 9-32 13-45
REFERENCES
Men 7-28 10-40
(1) TIETZ N.W. Text book of clinical chemistry, 3rd Ed. C.A. Burtis, E.R.
Ashwood, W.B. Saunders (1999) p. 652-657
Women 5-25 7-35
(2) Clinical Guide to Laboratory Test, 4th Ed., N.W. TIETZ (2006) p. 64-67
(3) YOUNG D.S., Effect of Drugs on Clinical laboratory Tests, 4th Ed. (1995) p.
Each laboratory should establish its own normal ranges for the 3-6 to 3-16.
population it serves. (4) HENRY R. J. andt al., Am J clin Path (1960), 34, 398
(5) Bergmeyer HU.,and al. Clin. Chem. (1978), 24, p.58-73
(6) IFCC Method for L-Alanine aminotransferase. J Clin. Chem., Clin.
PERFORMANCE CHARACTERISTICS Biochem.(1986), 24, p.481-495).
Within- Beetwen (7) MURRAY RL., « Alanine aminotransironase » in clinical chemistry. Theory,
run Normal level High level run Normal level High level analysis, and correlation.Kapan LA, Pesce AJ, (Eds), CV Mosby St Louis
N = 30 N = 33 (1984): 1090
Mean IU/L 32 141 Mean IU/L 39 98
S.D. IU/L 1.06 1.94 S.D.IU/L 1.15 1.45
C.V. % 3.3 1.4 C.V. % 2.9 1.5
Detection limit: approximately 7 IU/L
Sensitivity for 17 IU/L: approximately 0. 010 ∆Abs/min at 340 nm.
Comparison studies with commercially available reagent:
y = 0.9813 x – 0.6606 r = 0.9983
LINEARITY
The assay is linear up to 350 IU/L.
If ∆Abs./min > 0.200, reduce specimen volume or dilute specimen with
saline solution and reassay taking into account the dilution factor to
calculate the result. Linearity will depend on the specimen/reagent
ratio.
IVD REF LOT →
Manufacturer Use by In vitro diagnostic Temperature limitation Catalogue number See insert Batch number Store away from light sufficient for dilute with
Made in France Latest revision: www.biolabo.fr Revision: 18/12/2014
