Denaturing high-performance liquid chromatography technique platform applied to screen G6PD deficient variants

Area of Science:

• Clinical diagnostics
• Biochemical genetics
• Molecular biology

Background:

• Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common genetic disorder.
• Accurate screening for G6PD deficient variants is crucial for clinical management.
• Existing screening methods have limitations in sensitivity, specificity, or cost.

Purpose of the Study:

• To develop and evaluate a denaturing high-performance liquid chromatography (DHPLC) platform for screening G6PD deficient variants.
• To compare the performance of DHPLC with established methods like ARMS and DNA sequencing.
• To assess the accuracy, sensitivity, efficiency, and cost-effectiveness of DHPLC.

Main Methods:

• A DHPLC technique platform was developed for screening G6PD deficient variants.
• DHPLC was applied to 124 patients from diverse regions in China.
• Performance was compared against amplification refractory mutation system (ARMS) and DNA sequencing.

Main Results:

• DHPLC accurately identified known G6PD deficient variants, characterized by sharp peaks and verified by DNA sequencing.
• The method established a database of common G6PD variants in Chinese populations.
• DHPLC detected G6PD variants in negative controls and identified 34 heterozygous females, demonstrating high sensitivity and accuracy compared to ARMS, which showed false positives and negatives.

Conclusions:

• DHPLC is a highly advantageous method for screening G6PD deficient variants due to its accuracy, convenience, automation, and lower cost.
• DHPLC significantly aids in identifying female heterozygotes and individuals with G6PD deficiency clinical type IV.
• The developed DHPLC platform offers a superior alternative for G6PD deficiency screening.