Detection Of Two Common Glucose-6-phosphate Dehydrogenase Gene Point Mutations In Chinese By Single-tube Multiplex Polymerase Chain Reaction

Objective: To establish a rapid and simple gene diagnosis technique based on polymerase chain reaction (PCR) for detecting the most two common glucose-6-phosphate dehydrogenase(G6PD)gene mutations in Chinese, namely G1376T (G6PD Canton) and G1388A(G6PD Kaiping) and to apply it for screening the patients with G6PD deficiency in Guangxi. Methods: 213 samples with glucose-6-phosphate dehydrogenase deficiency and 20 males with normal G6PD activity were selected randomly from those sent to the pediatrics laboratory of Guangxi Medical University. Those samples were detected by the single-tube multiplex polymerase chain reaction and amplification refractory mutation system(ARMS) and the natural primers or mis-matched primers mediated PCR followed by restriction endomuclease analysis. Results got by the two methods were then compared to those got by the PCR followed by restriction endomuclease analysis method for concordance, accuracy, sensitivity and specificity. Two cases of G1376T and two cases of G1388A were selected randomly from those positive samples which were detected by single-tube multiplex polymerase chain reaction. They were identified by sequence analysis. Results: The single-tube multiplex PCR can detect G6PD gene G1376T and G1388A mutations at one polymerase chain reaction. Among 213 cases G6PD deficiency samples, 45 cases were G1376T, accounting for 21.13%. 83 cases were G1388A, accounting for 38. 97%. The two common G6PD genotypes added up to 128 cases, accounting for 60.09%. The results were the same as those being detected by ARMS. After the samples which had been detected by single-tube multiplex polymerase chain reaction were identified by PCR followed by restriction endomuclease analysis and by sequence analysis, we can find that there weren't false positive and false negative. Conclusions: The single-tube multiplex PCR for detecting G6PD G1376T and G1388A genotypes is accurate and sensitive. It is simple and rapid to practice for detecting the two common G6PD genotypes. It can be used as a routine method for screening G6PD genotypes in large scale.