孕妇血浆中胎儿DNA的提取及STR基因的检测

Extraction of fetal DNA in maternal plasma and detection of short tandem repeat gene

目的探索在孕妇血浆中提取胎儿DNA及STR基因的检测方法。方法提取10名健康孕妇(12～40周)血浆标本中胎儿游离DNA,对15个短串联重复序列(shorttandemrepeat,STR)基因位点和1个性别Amelogenin基因进行聚合酶链反应(polymerasechainreaction,PCR)扩增。同时,检测孕妇及其丈夫的基因组DNA,进行对照研究。结果经性别Amelogenin基因的检测,与胎儿实际的性别符合率为100%。经STR-PCR检测发现,所有10例孕妇血浆中均检测出父源性胎儿DNA的存在,检测数量为2.89±1.62。结论本实验采取的方法准确性较高,可作为符合国家法律规定范围内的一种无创性的产前诊断方法 。

Objective To explore the methods of extracting fetal DNA in maternal plasma and detecting STR gene. Methods The DNA templates was extracted by QIAamp Circulation Nucleic Acid Kit from 10 maternal（ 12 -40weeks）plasma. Multiplex polymerase chain reaction was amplified at ffteen different short tandem repeat （STR） loci and one Amelogenin gene. At the same time, testing pregnant women and their husbands genomic DNA, for control study. The results were confirmed by examination of newborns after delivery. Results The concordance rate between real fetal sex and sex determined by amplification of the Amelogenin gene was 100% （10/10）. The paternally inherited fetal alleles were detected by STR detection in all cases. Conclusion In this study, the approaches taken in the higher accuracy can be used as non- invasive prenatal diagnosis methods.