CLINICAL SIGNIFICANCE
Increased levels are observed in various
conditions of renal impairment. Gout,
Leukemia, Toxemia of pregnancy, Broncho
and lobular pneumonia and during cortisone
treatment.
Low values may be observed in Wilson's
disease and Fanconi's syndrome.
PRINCIPLE |
|
| UricAcid + O2 |
Uricase Allantoin + |
+2H2O |
CO2 + H2O2 |
2H2O2+DHBSAPODQuinoneimineDye |
+APP |
+4H2O |
The absorbance at 520nM is directly
proportional to uricacid concentration. |
REAGENTS SUPPLIED |
| Cat.No.A200 PACK SIZE: 15x1.2 mL |
| 1.Enzyme (1.2 mL) |
15 Vials |
| 2.Buffer(25 mL) |
1 Bottle |
| 3.Standard 5.0 mGs/dL (3 mL |
1 Vial |
| CATNO.A201 PACK SIZE: 10x10 Vials |
| 1.Enzyme (10mL) |
10 Vials |
| 2.Buffer(10mL) |
10Vials |
| 3.Standard 5.0 mGs/dL (3 mL) |
1 Vial |
| CAT NO. A 202 PACKSIZE: 10x10 Vials |
| 1.Enzyme (10mL) |
10 Vials |
| 2.Buffer(10mL |
10Vials |
| 3.Standard 5.0 mGs/dL (3 mL) |
1 Vial |
| COMPOSITION |
| Final concentration in the reactive medium: |
| Phosphates Buffer pH 7.7 |
50mMol/L |
| Amino-4-antipyrine |
0.6mMol/L |
| Peroxides |
>1000U/L |
| DHBSA |
>300U/L |
| Detergent |
Trace |
STANDARDS
Uricacid (297 µMol/L) 5.0 mGs/dL
Preparation of Working Reagent
1.Dissolve the substrate (Reagent No. 1)
with buffer (Reagent No.2) 1.2 mL in the case
ofCat.No.A200.
And 10 mL in the case of Cat. No. A 201 & A
202.
A uniform solution may take place after 5
minutes which is ready to use.
2.Markthedateofreconstitution.
STORAGE&STABILITY
Unopened reagent is stable for 18 months when
stored at 2-8°C.
The reconstituted reagent is stable for at least 2
weeks at 2-8°C away from light. Slight pink colour
in blankdoes not affect the test performance.
SAMPLE
Sample can be serum or plasma which has no sign of haemolysis. Uric acid is affected by the food intake containing nucleoproteins. Therefore, keep the patient fasting for at least 8 hours prior to sample collection. common anticoagulants have no interference on this assay. All samples should be handled as potential infective agents as no laboratory methods make conclusive finding for its safety. Therefore, adequate laboratory measures should be taken while handling such materials.
|
MANUAL METHOD
1.
| Pipette into 3 Test Tubes |
Blank mL |
Standrd mL |
Test |
| Working Reagent 1 |
1.0 |
1.0 |
1.0 |
| Standard (Reagent 3) |
- |
0.1 |
- |
| Serum/Plasma |
- |
- |
0.1 |
2.Mix well Incubate at 370C for 10 minutes.
3.Read at 520 nM (490 - 540 nM) or GREEN filter against blank.
For larger cuvette size add 2mL distilled water and read. The final colour is stable for 30 mins away from bright light.
Micro Method for discrete analysers
| ForCuvette Volume |
1.0mL |
0.5mL |
| Working Reagent |
1000 µL |
500 µL |
| Sample or Standard |
50µL |
20 µL |
Mix well. Incubate at 370C for 10 minutes. Read at 520 nM (490-540nM) against blank. NOTE : Programme the analyser using system parameters. A specific programme data sheet may be provided for each analyser upon request.
SYSTEM PARAMETERS
| Reaction |
= |
End-Point |
| Temperature |
= |
37°C |
| Wavelength |
= |
520 + 20nM |
| Standard Concentration |
= |
5.0mGs/dL |
| Absorbance Range |
= |
0-2°A |
| Reagent Volume |
= |
100 µL |
500 µL |
| Sample Volume |
= |
50µL |
20µL |
| Reaction Time |
= |
10Mins |
| Linearity |
= |
20.0mGs/dL |
| Max. limit of blank rgt. |
= |
0.1 °A |
| Final ColourStability |
= |
30Mins |
RESULTS
Compute
O.D. Test
UrciAcidinmGs/dL = --------------- x 5
O.D.STD
EXPECTED VALUES
Females: 2.7-6.5mGs/dL(160-387µMol/L) Males: 4.0-7.2 mGs/dL (237-428 µMol/L) 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 highly specific to various compounds. In vitro addition or contamination of Urates, Hydrogen Peroxide, Hypochlorate formaldehyde or any such compounds totally alter the results. This assay is linear to 20.0 mGs/dLofUricAcid.
For values higher than 20.0 mGs/dL repeat the test with serum diluted in 0.9% sodium chloride solution. Multiply the results by the dilution factor applied i.e. multiply by 2fora 1:1 dilution.
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, eleanliness of glasswares and accuracy of pippetting. It is appropriate to establish each laboratory's accuracy constant and interpret values. Similarly, laboratory findings should be established by clinical manifestations.
WARNING
This reagent system is for in vitro 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, orconsulta physician.
BIBLIOGRAPHY
1. Belis,M.E.Adv.Clin.Chem., 18(213)1976
2. Haeckel R.J., Clin. Chem. Biochem., 3 (14)
1976.
3. Henry R.J., Common C., Wrinkelman J.W.Eds.
Clinical Chemistry. Principle and Techniques.
Harper&Row, Hagerstown, MD1974.
4.TrinderP., J. Clin. Pathol., 22 (246) 1949.
5.Trivedi R., Berta E. and Reber L., Clin. Chem.
22(1223)1976.
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