用不提取核酸PCR扩增孕妇血浆中胎儿游离DNA的SRY和FMR1基因  

Extraction-free PCR for amplification of SRY and FMR1gene from free fetal DNA in maternal plasma

作　　者：陈斌[1] 严伟[1] 张琼[1] 邹晓[2] 周小棉[1] 钟志敏[1] 

机构地区：[1]广州医学院附属广州市第一人民医院检验科,广州510180 [2]广州医学院附属广州市第一人民医院妇产科,广州510180

出　　处：《临床检验杂志》2013年第4期253-255,共3页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金项目(81201163;81271730);广东省科技项目(2010B080702018);广州市医药卫生科技项目(201102A212028;20121A011035)

摘　　要：目的建立一种不提取核酸的孕妇血浆中胎儿游离DNA的检测方法。方法构建不提取核酸PCR法,用巢式PCR扩增Y染色体上的SRY基因序列和X染色体上的FMR1基因序列,检测44例孕妇血浆中游离DNA。根据随访判断结果的准确性。结果 44例孕妇经随访确认24例有男性胎儿,20例有女性胎儿。24例孕男性胎儿的孕妇血浆标本中有21例扩增出SRY基因和FMR1基因,敏感性为87.5%(21/24);20例孕女性胎儿的孕妇血浆标本中有17例扩增出FMR1基因,敏感性为85.0%(17/20)。结论建立的不提取核酸PCR法具有快速、简便等优点,可用于性连锁遗传病的产前诊断。Objective To develop a rapid and simple method for the detection of free fetal DNA in maternal plasma. Methods A DNA extraction-free PCR method was developed. Both the gene-speeific sequences of FMR1 on X chromosome and SRY on Y chromo- some were amplified simultaneously by multiplex nested PCR. The free fetal DNA in maternal plasma from 44 pregnant women was de- tected. On the basis of follow-up results the accuracy of the developed direct PCR method was evaluated. Results Both the FMR1 and SRY gene-specific sequences were detectable in 21 of 24 maternal plasma samples obtained from the pregnant women carrying male fe- tus. Single FMR1 gene-specific sequence was detectable in 17 of 20 plasma samples of pregnant women carrying female fetus. The ac- curacy rate of the method for gender prediction of fetus were 87.5% （21/24） for male fetus and 85% （17/20） for female fetus respec- tively compared with the gender identification after delivery. Conclusion The DNA extraction-free PCR developed in this study was rapid, simple for detection of free DNA in maternal plasma, thus it should be useful for prenatal diagnosis of sex-linked genetic disease.

关 键 词：胎儿游离DNA SRY基因 FMR1基因 不提取核酸聚合酶链反应 

分 类 号：R714.55[医药卫生—妇产科学]