中国人常见G6PD基因突变的快速检测

目的葡萄糖-6-磷酸脱氢酶（glucose-6-phosphate dehydrogenase,G6PD）缺乏症是世界上最常见的遗传性红细胞酶缺陷病。本文探讨建立一种简便快速、准确可靠、经济实用的G6PD缺乏症分子诊断技术。方法采用DNA测序技术通过双盲法对所建立的单管多重PCR结合分子杂交技术进行方法学评价。结果所建立的新技术可成功地检出上述中国人常见的G6PD基因突变并能清楚区分男性杂合子、女性杂合子或纯合子或双重杂合子。盲法分析结果显示,该技术对经DNA直接测序确诊的G6PD标本的诊断准确度和重复性均达到100%。结论单管多重PCR结合分子杂交技术可准确、快速地检测中国人常见的G6PD基因突变且具有经济和实用的优点,非常适合于G6PD缺乏症的大人群分子筛查和临床样品的基因诊断。

Rapid detection of common G6PD gene mutation in Chinese

Objective Glucose-6-phosphate dehydrogenase（G6PD） deficiency is a worldwide common inherited hemolytic disorder with enzyme abnormality in red blood cell. This study was to develop a simple, rapid, accurate and cost-effective molecular assay for G6PD gene mutation detection. Methods The results could be interpreted by the color of hybridization spot after washing and coloring membrance. Direct DNA sequencing was used to evaluate the established STMPCR combined with molecular hybridization in a double-blind study. Results The established RDB was able to detect successfully the six common mutations among Chinese and distinguish clearly every genotype of G6PD including hemizygotes in male, heterozygotes and homozygotes as well as compound heterozygotes in female. Double-blind study showed that the molecular hybridization assay for G6PD gene genotyping was in complete concordance with the direct DNA sequencing and had very good repeatability. Conclusion The established STMPCR combined with RDB could rapidly and accurately detect common G6PD gene mutations among Chinese with the advantages of cost-effectiveness and practicability. It is very suitable for large-scale population screening and rapid clinical molecular genotyping in G6PD deficiency.