两重巢式PCR检测母体外周血中胎儿游离DNA  

Detection of Free Fetal DNA from Maternal Blood by Multiplex Nested PCR

作　　者：袁媛[1] 陈思礼[2] 周新[1] 

机构地区：[1]武汉大学第二临床学院基因诊断中心,武汉430071 [2]中南民族大学生命科学学院,武汉430074

出　　处：《中南民族大学学报（自然科学版）》2008年第4期44-46,共3页Journal of South-Central University for Nationalities：Natural Science Edition

基　　金：国家自然科学基金资助项目(30672012);湖北省杰出青年基金资助项目(2006ABB009)

摘　　要：使用两重巢式PCR技术检测母体外周血中胎儿游离DNA，为非损伤性产前鉴定胎儿性别和诊断单基因疾病奠定基础．采用改良的chelex100方法提取60例孕龄为16～36周孕妇的外周血浆中游离胎儿DNA，依据NCBIX染色体长臂ATL1位点，Y染色体上DYS14位点序列，设计并合成引物，用巢式PCR同时扩增2个基因片段，以鉴定胎儿性别．结果表明；有ATL1位点和DYS14位点的扩增产物261bp和198bp鉴定为男性胎儿，而仅有261bp扩增产物鉴定为女性胎儿．均以真实出生性别进行确认后发现，此方法能有效提高检测特异性．两重巢式PCR能够简便并准确地鉴定胎儿性别，对单基因疾病特别是X-连锁遗传病的诊断有积极意义．To determine fetal gender and to use it for single gene genetic prenatal diagnosis, the multiplex nested PCR was obtained. Fetal DNA in peripheral blood from pregnant women aged from 16 weeks to 36 weeks was extracted by chelex 100. The sequences of ATL1 located in X chromosome and DYS14 located in Y chromosome from NCBI and the nested PCR primers were designed and synthesized. The gender was determined by the nested PCR with the two located genes applified simultaneously. The amplicons of 261bp and 198 bp proved that both ATL1 and DYS14 were amplified, identifying the fetus was male. Conversely, the female fetus was only 261bp amplicon. The gender determination was compared with the real gender after birth, and this detection can increase the specificity. It is significant and beneficial to determining the fetal gender and diagnosing the single gene diseases, especially for X-linked genetic diseases before birth with the peripheral blood from preganant women by multiplex nested PCR.

关 键 词：母体外周血 胎儿游离DNA 两重巢式PCR 性别鉴定 单基因疾病 

分 类 号：Q789[生物学—分子生物学]