What are the principles of the following tests?
a. Fluorescent spot test
b. Dye reduction test
c. Ascorbate cyanide test
d. Rate reduction test
What will be an ideal response?
Answer: Fluorescent spot test: Whole blood is added to a mixture of G6P, NADP, and saponin. G6PD enzyme metabolizes G6P and produces NADPH (which fluoresces). NADP does not. Lack of fluorescence indicates G6PD deficiency.
Dye reduction test: similar to fluorescent spot test except for indicator: blue color to colorless presence of G6PD; blue color remains absence of G6PD
Ascorbate cyanide test: nonspecific test. Tests the ability of RBC catalase to inhibit hydrogen peroxide formation. G6PD-deficient patients allow hydrogen peroxide to form, which promotes the formation of methemoglobin. The presence of methemoglobin turns the solution brown.
Rate reduction test: Measures rate of conversion of NADP to NADPH via spectrophotometry at 340 nm.
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