ACCUCARETM
G-6PD
(Qualitative Screening Method)
Catalog No: Product Description: Pack Size:
G-6PD 10 G-6PD Screening test 10 x 1 test

CLINICAL SIGNIFICANCE OF G6PD TEST

G6PD deficiency is the underlying cause of certain drug induced hemolytic anaemia.

PRINCIPLE
Glucose- 6 phosphate dehydrogenase, present in the red cell hemolysate, acts on Glucose- 6 phosphate and reduces NADP which, in the presence of PMS, reduces the blue colored 2,6 Dichlorophenolindophenol into a colorless form leaving behind the original cherry red color of the hemolysate. The rate of decolorisation is proportional to the enzyme activity.

SAMPLE
Whole blood collected in E.D.T.A bulb.

REAGENTS
1. SUBSTRATE VIALS
2. BUFFER, Ph 8.5
3. MINERAL OIL

REAGENT STORAGE & STABILITY
Reagent 1 & 2 are to be stored at 2-8 °C.

PRECAUTIONS
Use fresh blood samples, as enzyme activity reduces on refrigeration. do not use Heparin as an anticoagulant, Heparin interferes with the results.

WORKING REAGENT PREPARATION
Tap the subtrate vials gently on a flat surface to dislodge all the substrate powder. Just before use, reconstitute each substrate vial with 0.5 ml of Buffer Reagent. Gently swirl to dissolve and allow to stand for five minutes.

PREPARATION OF RED CELL HEMOLYSATE
Enclosed below is table showing amount of blood required to prepare the hemolysate corresponding to the Hemoglobin concentration (G/DL).

REFERENCES:
S.K.Sood et al., The Indian Journal of Path. And Micro., 24
(1981),89. Lubin, B.H. and Oski, F.A., J. Pediatr. 70 (1967)
Hemoglobin Cone. GM/DL Quantity of Blood (ml)
7.0 to 9.50 0.04 ml
9.60 to 11.50 0.03 ml
11.6 to 13.50 0.025 ml
13.6to 15.0 0.02ml
In 1.0 ml of Distilled water add 20 ml (0.02 ml) of well mixed E.D.T.A. whole blood sample. Mix well & allow to stand for 5 minutes at room temperature.

ASSAY PROCEDURE
1. Add 1 ml of the hemolysate to the reconstituted substrate vial & mix gently by swirling.
2. Add immediately 1 ml of the mineral oil.
3. Replace the plug & the cap tightly. Incubate undisturbed
at 37°C for 60 minutes in dark.

NOTE : The vials should be incubated undisturbed as the
introduction of air from the atmosphere will allow the blue color to reappear & will give erroneous results.

INTERPRETATION OF RESULTS

DECOLORISATION TIME

Normal Subjects: Upto 60 minutes.

G6PD Deficient subjects :
(In Heterozygous males & Homozygous Females) Decolorisation time is 2 to 24 hrs. In Heterozygous females who are carriers the cell population is mixed with normal & deficient cells. The distribution of deficient cells varies from individual to individual ranging from 20% to 80%. Hence some such subjects may give results overlapping over normal as well as abnormal time specifications i.e. the Decolorisation in some heterozygotes will be between 5 to 20 minutes (normal) & for some heterozygotes the same wilt be 2 hrs, or more.

NOTES
1. Samples having hemoglobin content below 15 GM%, proportionately use more amount of blood for preparation of the hemolysate.
2. Blood with high reticulocyte count may give false normal results even though the patient is enzyme deficient as reticulocytes generally have a higher G6PD activity than adult red cells.
Lab Care Diagnostics (India) Pvt. Ltd.
16-A, 'A' Wing , 1st Floor Sita Estate Mahul RoadAziz Baug, Chembur Mumbai 400 074. India
Phone: 00 91 22 2554 2109/ 00 91 22 2554 1558 Fax: 00 91 22 2554 3541
E-mail : accucare@labcarediagnostics.com