Yang Linlin, Lv Qiaoying, Chen Wei, Sun Jian, Wu Yu, Wang Yiying, Chen Xiong, Chen Xiaojun, Zhang Zhenbo
The Reproductive Medicine Center of Department of Obstetrics and Gynecology, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 200080, China.
Department of Obstetrics and Gynecology, Shanghai First People's Hospital, Baoshan Branch, Shanghai 201900, China.
Oncotarget. 2017 Jun 29;8(40):67805-67809. doi: 10.18632/oncotarget.18852. eCollection 2017 Sep 15.
Preimplantation genetic diagnosis (PGD) has successfully assisted couples with genetic diseases to conceive healthy babies during the past decades. However, biopsy of the blastomere has potential lesion to the embryos which commonly results in abortion. Thus, a noninvasive PGD is needed. In the past, the presence of genetic materials in maternal plasma or serum has triggered a great innovation of noninvasive prenatal diagnosis. Nevertheless, it is not clear whether embryonic DNA is also present in embryonic culture medium. Here, a rapid-boiling method has been used to harvest DNA from the medium or the discarded embryos, following Polymerase Chain Reaction (PCR) was applied to detect the dissociative DNA by amplifying SRY gene (Y-chromosome). For the first time, the Y sequences were detected in the medium which were used to culture embryo for above 3 days. None of the positive signal was examined in Day 1 and Day 2 embryonic culture medium. Our findings suggest that the Y chromosome fragments from the embryo may release into its culture medium. If validated in a larger cohort, detection of SRY gene may prove to be a useful method to screen Y-linked genetic disease. More importantly, detecting the free DNA in the embryonic culture medium may represent a novel strategy for noninvasive PGD.
在过去几十年里，植入前基因诊断（PGD）已成功帮助患有遗传疾病的夫妇孕育健康婴儿。然而，对卵裂球进行活检会对胚胎造成潜在损伤，这通常会导致流产。因此，需要一种非侵入性的PGD。过去，母体血浆或血清中遗传物质的存在引发了非侵入性产前诊断的重大创新。然而，尚不清楚胚胎培养基中是否也存在胚胎DNA。在此，采用快速煮沸法从培养基或废弃胚胎中提取DNA，随后应用聚合酶链反应（PCR）通过扩增SRY基因（Y染色体）来检测解离的DNA。首次在用于培养胚胎3天以上的培养基中检测到Y序列。在第1天和第2天的胚胎培养基中未检测到任何阳性信号。我们的研究结果表明，来自胚胎的Y染色体片段可能释放到其培养基中。如果在更大的队列中得到验证，检测SRY基因可能被证明是筛查Y连锁遗传病的一种有用方法。更重要的是，检测胚胎培养基中的游离DNA可能代表一种非侵入性PGD的新策略。
