摘要
目的探讨微滴式数字PCR(droplet digital PCR,ddPCR)技术在单基因遗传病无创产前诊断中的应用。方法应用ddPCR技术对母体外周血中胎儿游离核酸、羊水DNA进行父源性致病基因位点检测,应用Sanger测序技术对羊水DNA进行父源性致病基因位点确认。结果ddPCR技术对孕妇外周血游离核酸检测结果与羊水DNASanger测序结果一致,显示两家系胎儿均携带父源性致病基因突变位点。结论ddPCR技术可对微量样本进行准确、灵敏、稳定的定性分析,检测孕妇外周血中胎儿游离核酸,为父源性单基因遗传病产前诊断提供了有效、快捷、安全的检测方法。
Objective To assess the value of droplet digital PCR (ddPCR) for non-invasive prenatal diagnosis of single gene disease in two families. Methods Paternal mutation in celbfree DNA derived from the maternal blood and amniotic fluid DNA was detected by ddPCR. Suspected mutation in the amniotic fluid DNA was verified with Sanger sequencing. Reverse The result of ddPCR and Sanger sequencing indicated that the fetuses have carried pathogenic mutations from the paternal side in both families. Conclusion Droplet digital PCR can accurately detect paternal mutation carried by the fetus, and it is sensitive and reliable for analyzing trace samples. This method may be applied for the diagnosis of single gene diseases caused by paternal mutation using peripheral blood sample derived from the mother.
作者
徐佩文
邹洋
李杰
黄色新
高明
康冉冉
解洪强
王丽娟
颜军昊
高媛
Xu Peiwen , Zou Yang , Li Jie , Huang Sexin , Gao Ming , Kang Ranran , Xie Hongqiang , Wang Lijuan , Yan Junhao, Gao Yuan(Center for Reproductive Medicine, Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, The Key Laboratory for Reproductive Endocrinology of Ministry of Education, J inan , Shandong 250012, Chin)
出处
《中华医学遗传学杂志》
CAS
CSCD
2018年第2期224-227,共4页
Chinese Journal of Medical Genetics
基金
山东省重点研发计划(2016GGH4522)