CLINICAL SIGNIFICANCE
A high activity of ALT is the clinical indication of Hepatitis and or liver cell damage.
PRINCIPLE
L - Alanine ALT Pyruvate
-------------® +
a - ketoglutarate L-glutamate
Pyruvate LDH Lactate
+ -----------® +
NADH NAD
The catalytic activity of ALT is measured by the rate of NADH dissociated to form NAD at 334, 340 or 365 nM which is directly proportional to the ALT units.
COMPOSITION |
| Tris buffer |
100mMol/L |
| a - ketoglutarate |
15 mMol/L |
| L - aspartate |
500 mMol/L |
| LDH |
> 1200 U/L |
| NADH |
0.24 mMol/L |
| L-alanine |
500mMol/L |
Contains stabilisers, preservatives and non-reactive fillers. |
| REAGENTS SUPPLIED |
| I.Cat. No. LS365 |
3x10mL |
| Reagent 1 Buffer |
3 x 9 mL |
| Reagent 2 NADH |
3 x 1 mL |
| 2.Cat. No. LS 366 |
10x10mL |
| Reagent 1 Buffer |
10x9 mL |
| Reagent 2 NADH |
10x1 mL |
| 3.Cat. No. LS 367 |
4x50 mi- |
| Reagent 1 Buffer |
4 x 45 mL |
| Reagent 2 NADH |
4 x 5 mL |
Preparation of Working Reagent
Mix R2 to R1.
STABILITY OF REAGENT
Unopened reagent is stable until expiry date marked on each label. Working reagent Is stable for 30 days, when stored at 2-8°C & protected from light.
SAMPLE
Use fresh serum or heparinised (or EDTA Na+) plasma which shows no sign of haemolysis. AsAT activity reduces at a rate of 8%/hour at 2-8°Cand10%/hourat30°C.
Avoid fluoride plasma.
|
METHOD FOR AUTOANALYSERS
| Temperature |
37°C |
| Working Reagent |
500 µL |
| Sample |
50 µL |
| Factor |
1746 |
Mix well. Take one minute reading at 340 nM after a delay of 60 seconds at 37°C.
NOTE: Programme the analyser using system parameters. A specific programme data sheet may be provided for each analyser upon request. Reagent, sample volume and factor can be altered proportionately.
SYSTEM PARAMETERS
| Reaction |
= UV-Kinetic |
| Direction of Reaction |
= Inverse |
| Temperature |
= 37°C |
| Wavelength |
= 340 nM |
| Factor % |
= 1746 |
| Absorbance Range |
= 0-2°A |
| Cuvette Path Length |
= 1 cm |
| Delay Time Seconds |
= 60 seconds |
| Interval Seconds |
= 20 seconds |
| No. of Readings |
= 3 |
| Linearity |
= 300 U/L |
| Blank |
= DW |
| Reagent Volume |
= 500 µL |
| Sample Volume |
= 50µL |
RESULTS
Compute average rate of absorbance per minute x 1746.
EXPECTED VALUES
| TEMPERATURE |
37°C |
| MEN IU/L |
0-40 |
| WOMEN IU/L |
0-38 |
As with all diagnostic methods, the final diagnosis should not be made on the result of a single test as well as laboratory diagnosis must be confirmed with clinical manifestations. LIMITATIONS
This reagent system is linear up to
300 U/L.
If the test exceeds the linearity limit dilute the sample with 0.9% Sodium Chloride and rerun the test and multiply the result with dilution factor. This methodology highly depends on accuracy and precision of instrument and professionals.
QUALITY CONTROL
To ensure adequate quality control, each kit should be tested against a standard control sera. It should be realised that the use of quality control material checks both instrument and reagent function together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glasswares and accuracy of pipetting.
It is appropriate to establish each laboratory's accuracy constant and interpret values accordingly. Similarly, laboratory findings should be established by clinical manifestations.
WARNING
This reagent system is for invitro Diagnostic use only. This reagent system is containing preservatives and components that have not established for safety if contacted on broken skin or eye or taken orally. In case of such incidents wash off with plenty of water, or consult a physician.
BIBLIOGRAPHY
- Jung K. Fechner C, Egger E, J. Cli. Biochem(1976)14,53.
- Thefeld W, et. al., Dtsch. Med. Wschr. (1974)99,343.
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