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G6PDH Qualitative Dye 12

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G6PDH Qualitative Dye 12
G6PDH Qualitative Dye 12
Price And Quantity
  • Kit/Kits
  • 1
  • Kit/Kits
  • 900 INR
    Product Specifications
    • other
    • Biochemistry reagent
    • 18 Months
    • Other
    • Auto Zyme NEW G6PDH (Qualitative-Dye) is a reagent set with simple procedure and easy technique. Auto Zyme NEW G6PDH (Qualitative-Dye) is available in convenient pack-size of 12 tests.
    • Biochemistry reagent
    • Liquid
    • Other
    • Reagents
    • Other
    • ACC776556
    • Industrial
    • Industrial Lab Chemicals
    • 99%
      Product Description
      INTRODUCTION
      • Auto Zyme NEW G6PDH (Qualitative-Dye) is a reagent set for qualitative determination (screening) of G6PDH deficiency in erythrocytes by decolourisation method.
      • Auto Zyme NEW G6PDH (Qualitative-Dye) is a reagent set with simple procedure and easy technique.
      • Auto Zyme NEW G6PDH (Qualitative-Dye) is available in convenient pack-size of 12 tests.
      PRINCIPLE

      Glucose-6-Phosphate Dehydrogenase (G6PDH) present in haemolysate acts on substrates, Glucose-6-Phosphate and NADP, giving NADPH, which in presence of PMS (Phenazine methosulfate) decolorises blue coloured indophenol dye (DCPIP) leaving behind colour only due to haemolysate. The rate of reaction is proportional to G6PDH present, hence time required for decolorisation is inversely proportional to G6PDH activity in the haemolysate.

      PREPARATION OF WORKING REAGENT

      Reconstitute one vial of R1 (Co-enzyme/Substrate) with 0.5 ml R2 (Buffer). Mix well to allow complete dissolution and use within 5 minutes.

      REAGENT STORAGE & STABILITY

       

      • Freshly prepared working reagent should be used.
      • The reagent kit should be stored at 2 - 8 degree C & is stable till expiry date indicated on the label. DO NOT FREEZE THE REAGENT.
      REAGENT COMPONENTS

       

      R1

      Co-enzyme/Substrate

      13 vials

      R2

       Buffer

      1 bottle

      R3

      Lysing Reagent

      1 bottle

      R4

      Inert oil

      1 bottle

       SPECIMEN COLLECTION  & PRESERVATION

      • Whole blood should be collected in a clean dry container using EDTA as an anticoagulant. Heparin should not be used as it interferes with the reaction.
      • Finger prick blood may be used provided the hemoglobin content is close to 15 gm/dl.
      • For an unknown sample the hemoglobin content must first be estimated and aliquot of blood may be corrected for low hemoglobin content
      PROCEDURE

      Estimate Hemoglobin content (gm/dl) of whole blood. If the hemoglobin content is significantly less than 15 gm/d1, adjust the hemoglobin content by proportionately increasing the aliquot of whole blood during preparation of red cell hemolysate.

      PREPARATION OF RED CELL  HAEMOLYSATE

      Given below is a table showing quantity of blood required for 1 ml lysing reagent corresponding to the hemoglobin concentration Hb gm/dl.

      Hemoglobin Concentration
      (gm/dl)

      Quantity of blood to be taken
      (ml)

      7.0-9.5

      0.04

      9.5-11.5

      0.03

      11.6-13.5

      0.025

      13.6-15.0

      0.02

      STEP 1 : HAEMOLYSATE PREPARATION :

      R3 (Pre-cooled lysing reagent) Fresh whole blood: 1.0 ml

      0.02 ml

      Mix well and keep it in refrigerator (2 - 8 degree C ) for 10 - 15 minutes and use as given below :

      STEP 2 :
      • Transfer completely the red cell haemolysate to the freshly prepared working reagent. Shake well.
      • Immediately overlay 1.0 ml of R4 (inert oil) on the reaction mixture.
      • Seal the vial tightly, using the plug and cap to make it air tight, incubate at 37 degree C.
      • Observe the change of initial blue to brownish colour. 
      INTERPRETATION 

      • In normal subjects, decolourisation time is between 30 -.60 minutes.
      •  In G6PDH deficient subjects, (heterozygous males and homozygous females) decolourisation time is between 2 to 24 hours.
      •  In heterozygous females, who are carriers, the cell population is mixed with normal and 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 decolourisation time in some heterozyous will be between 30 - 60 min. (normal) and for some heterozygous the same will be 2 hours or more.
      OBSERVATIONS
      • Observe the reaction mixture at 30 minutes for decolorization. If the decolorization is incomplete, observe every 5 minutes (or shorter intervals) thereafter until the decolorization is complete. If the decolorization takes longer time than 60 minutes, increase the interval time between observations and follow up for 4 - 8 hours or more.
      • In G6PDH deficiency the time taken for decolorization will exceed from 2 hours to 24 hours.
      PROCEDURE LIMITATIONS
      •  Sample may give false normal result in a deficient subject if the reticulocytes count is high, as reticulocytes have a higher G6PDH activity than adult red cells. This is of special importance if the test is carried out immediately after a haemolytic episodes in drug (primaquine or any such) sensitive subjects.
      • After initial 15 minutes it is better to observe the reaction tube at an interval of 5 minutes or less as some of the sample may reach the end point and then slowly turn blue again, due to re-oxidation of the dye.
      • Observation of the colour change should be restricted to the reaction mixture below the layer of oil and not at the interphase.
      • Vitamin C supplements or large amount of dietary intake of vitamin C may interfere with the reaction.
      QUALITY CONTROL
      • Always run one control sample of a known, healthy non-deficient subject on opening a kit.
      • Factors which might affect the performance of this test include temperature, cleanliness of glassware & accuracy of pipetting.




      Trade Information
      • EXW ABPL
      • Paypal, Letter of Credit at Sight (Sight L/C), Letter of Credit (L/C), Cheque, Cash in Advance (CID), Cash Against Delivery (CAD)
      • 10000 Per Day
      • 2 Days
      • Yes
      • packaging : Empity
      • Africa, Middle East, Western Europe, Eastern Europe, South America, North America, Central America, Australia, Asia
      • All India
      • certification : iso
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