应用三引物荧光PCR-Sanger测序法检测FMR1全突变者和前突变携带者  

Tri-primer-florescence PCR-Sanger sequencing method for screening of full and pre-mutations of FMR1 gene

作　　者：沙莎[1] 贺学[1] 袁东亚[1] 张建芳[2] 康龙丽[1] 

机构地区：[1]西藏民族大学医学部,陕西省咸阳市712082 [2]第四军医大学西京医院,西安710000

出　　处：《中华医学遗传学杂志》2016年第6期844-848,共5页Chinese Journal of Medical Genetics

基　　金：西藏自治区科技厅自然科学基金项目（12KJZRYMY09）;西藏民族学院重大项目培育计划（12myZP03）

摘　　要：目的检测脆性X综合征患者或携带者FMR1基因5′非编码区CGG序列的重复数。方法应用三引物荧光PCR-Sanger测序法检测一例男性脆性X综合征疑似患者和一名外表正常孕妇及胎儿羊水样本，并选择阳性对照和阴性对照进行可靠性验证。结果三引物荧光PCR-Sanger测序法可准确检出阴性及阳性对照的FMR1基因CGG的重复数。男性受检者CGG的重复数大于200，判断为全突变患者；孕妇CGG重复数为35和115，判断为杂合型前突变携带者；胎儿羊水CGG的重复数大于200，判断为男性全突变胎儿。经家属知情同意，对流产胎儿进行检测，证实了上述产前诊断结果。结论三引物荧光PCR-Sanger测序法可作为基层常规临床检测脆性x综合征患者／前突变携带者FMR1基因CGG重复数的快捷有效可靠方法，同时也适合在基层实验室用于大规模筛查前突变携带者候选人群。Objective To screen for CGG syndrome and carriers of pre-mutations. Methods repeats in the FMR1 gene among patients with fragile X Potential full and pre-mutations of the FMR1 gene were detected with a Tri-primer-floreseence PCR-Sanger sequencing method. The results were validated with positive and negative controls. Results All positive and negative controls were confirmed. A male patient was found to have〉 200 CGG repeats （full mutation）. For a pregnant women who was heterozygous for 35/115 CGG repeats, 〉 200 CGG repeats were also found with amniotic fluid sample from her fetus who was a male. The result was confirmed by following selective abortion with informed eonsent. Conclusion Tri-primer-florescence PCR-Sanger sequencing is a simple, effective and reliable method for routine screening of patients/carriers with full/pre-mutations of the FMR1 gene in the population.

关 键 词：脆性X综合征 三引物荧光PCR-Sanger测序 FMR1基因 前突变 全突变 

分 类 号：R4[医药卫生—临床医学]