检验医学与临床2016，Vol.13Issue(z1)：6-9,4.DOI:10.3969/j.issn.1672-9455.2016.25.003
G6PD缺乏症基因诊断方法的建立
Development of a new method for genotype of G6PD deficiency
摘要
Abstract
Objective To establish an effective ,feasible ,economic ,rapid and pollution‐free practical assay for genetic diagnosis of glucose‐6‐phosphate dehydrogenase(glucose‐6‐phosphate dehydrogenase ,G6PD)deficiency .Methods A multiple fluorescence PCR system based Taqman MGB probes was performed to amplify the target fragments of G 6PD gene ,and detect the domestic main 6 mutation sites kinds(1376G> T ,1388G>A ,95A>G ,871G>A ,392G> T ,1024C> T) .At the same time ,the beta actin gene moni‐tors the whole PCR system .Repeatability was tested in 30 samples with known genotype .A double‐blind trial for 200 unknown samples was performed by using multiplex fluorescence PCR assay and DNA sequencing simultaneously .Results The established multiple fluorescence PCR assay based Taqman MGB probes was able to detect the most common 6 mutations of G6PD gene simul‐taneously in 1 hour ,and both specificity and accuracy reached 100% .Among the 200 unknown samples ,34 mutational cases was de‐tected .All the genotyping results of multiple fluorescence PCR assay based Taqman MGB probes were in complete concordance with direct DNA sequencing ,and both specificity and accuracy reached 100% .Conclusion The established multiple fluorescence PCR assay based Taqman MGB probes ,detecting samples in 1 hour without open tubes after PCR ,is a rapid ,effective and feasible method of G6PD deficiency gene diagnosis .关键词
葡萄糖-6-磷酸脱氢酶/基因突变/多重Taqman MGB探针荧光PCR法/基因诊断Key words
glucose-6-phosphate dehydrogenase/gene mutation/multiple fluorescence PCR system based Taqman MGB probes/gene diagnosis引用本文复制引用
李印淑,刘晶晶,刘福平,刘梦莹,李维,刘丽益,张静..G6PD缺乏症基因诊断方法的建立[J].检验医学与临床,2016,13(z1):6-9,4.基金项目
深圳市南山区科技局项目（南研卫2014006）。 （）
