Principle
Glucose-6-Phosphate Dehydrogenase (G6PDH) present in hemolysate acts on substrates, Glucose-6-Phosphate and NAD, giving NADPH, which in presence of PMS (Phenazine methosulfate) decolorises blue coloured indophenol dye (DCPIP) leaving behind color only due to hemolysate.
The rate of reaction is proportional to G6PDH present, hence time required for decolorization is inversely proportional to G6PDH activity in the hemolysate
Procedure
Estimate Hemoglobin content (gm/di) of whole blood. f the
hemoglobin content is significantly less than 15 gm/dl, adjust
the hemoglobin content by proportionally increasing the
aliquot of whole blood during preparation of red cell hemolysate.
Quality Control
To ensure adequate quality control, it is recommended that each batch should include normal and an abnormal commercial reference control the serum. It should be realized that the use of quality control material checks both instrument and reagent functions together. Factors which might affect the performance of this test include proper instrument function, temperature
control, cleanliness of glassware and accuracy of pipetting.
Other Details
Freshly prepared working reagent should be used.
The reagent kit should be stored at 2- 8°C & is stable till expiry date
indicated on the label. DO NOT FREEZE THE REAGENT.
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