A novel cytofluorometric assay for the detection and quantification of glucose-6-phosphate dehydrogenase deficiency

Area of Science:

• Biochemistry
• Genetics
• Hematology

Background:

• Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a prevalent X-linked enzymopathy impacting millions globally.
• Current diagnostic methods like genetic and biochemical assays have limitations in cost-effectiveness and detecting female heterozygotes.
• Existing cytochemical assays, while capable, involve significant technical complexity and labor.

Purpose of the Study:

• To introduce a novel, simple cytofluorometric method for assessing G6PD deficiency at the single-erythrocyte level.
• To provide a robust, economical, and high-throughput alternative to existing diagnostic and screening techniques.
• To evaluate the concordance of the new method with established genetic and biochemical assays.

Main Methods:

• Development of a novel cytofluorometric assay based on the classic methemoglobin reduction test.
• Assessment of G6PD deficiency at the individual erythrocyte level using flow cytometry.
• Preliminary testing and validation in Malian children.

Main Results:

• The novel cytofluorometric method demonstrated strong concordance with established genetic and biochemical techniques in preliminary testing.
• The assay proved to be robust and economical.
• The method is suitable for both screening and research applications, including high-throughput analysis.

Conclusions:

• The developed cytofluorometric method offers a simple, accurate, and economical approach for diagnosing G6PD deficiency.
• This assay overcomes limitations of current methods, particularly for screening in populations with allelic heterogeneity and for detecting female heterozygotes.
• The technique is well-suited for high-throughput screening and research, potentially improving global G6PD deficiency management.