贵州医科大学学报

2011, v.36;No.148(01) 15-18+22

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孕妇血清中游离胎儿DNA的分离及鉴定
Isolation and Identification of Free Fetal DNA in Maternal Serum

罗衡丽;韦四喜;刘丽荣;赵鲁强;张小蕾;
LUO Hengli,WEI Sixi,LIU Lirong,ZHAO Luqiang,ZHANG Xiaolei (Department of Clinical Laboratory Science,Guiyang Medical College,Guiyang 550004,Guizhou,China)

摘要(Abstract):

目的:建立一种稳定的提取母体血清DNA和鉴定胎儿DNA的方法。方法:采用裂解液煮沸法提取孕早、中、后期共90例孕妇血清DNA,应用PCR技术及亲子鉴定原理检测孕妇血清及夫妇双方DYZ1基因、DXS6804、DXS6799和D18S51以确认母体血清中胎儿DNA。结果:男胎孕妇血清中DNA DYZ1基因早、中、晚期的检出率分别为75%、88.89%、100%;血清中胎儿DNA的DXS6804、DXS6799、D18S51的孕早、中、后期检出率分别是76.67%、93.33%、100%。结论:裂解液煮沸法能提取孕妇血清游离DNA,且简便、快捷、提取效率高,可用于后续产前诊断;孕妇血清中存在胎儿游离DNA,并随孕周的增加而明显增加;STR位点扩增及亲子鉴定方法的应用,弥补了DYZ1序列仅能用于鉴定男性胎儿DNA的不足,可作为一种鉴定胎儿DNA的更为可靠的方法;X-STR(DXS6804、DXS6799)和Y染色体上的DYZ1位点可用于胎儿性别的鉴定,为性连锁遗传病的早期筛选提供了一种新的方法。
Objective:To establish a stable method for isolation and identification of fetal DNA in maternal serum.Methods: Serum DNA of 90 pregnant women was isolated with boiling lysis method.PCR was used to detect DYZ1 gene,and short tandem repeat sequences(STR) of DXS6804,DXS6799 and D18S51 in serum of couple.Fetal DNA sequences were identified following paternity test principle.Results: Detecting rates of male fetal DYZ1 gene in early,intermediate and late stages were 75%,88.89%,and 100% respectively.Detecting rates of fetal DNA sequences of DXS6804,DXS6799,and D18S51 in early,intermediate and late stages were 76.67%,93.33%,and 100% respectively.Conclusions: Fetal DNA exists in maternal serum and its concentration increases significantly as gestational age increased.Free DNA in maternal serum could be easily and efficiently isolated with boiling lysis method and could be used in following prenatal diagnosis.STR loci amplification and application of the paternity testing methods cover the shortcomings of DYZ1 gene detectation that is useful only in identification of male fetal DNA.They are more reliable for identification of fetal DNA.X-STR(DXS6804,DXS6799) and DYZ1 gene on Y chromosome could be used for identification of fetal sex,which provides a new approach for early screening of sex-linked genetic diseases.

关键词(KeyWords): 孕妇;胎儿;基因;DNA;血清
pregnant women;fetus;genes;DNA;serum

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基金项目(Foundation): 贵州省教育厅自然科学研究基金资助项目

作者(Author): 罗衡丽;韦四喜;刘丽荣;赵鲁强;张小蕾;
LUO Hengli,WEI Sixi,LIU Lirong,ZHAO Luqiang,ZHANG Xiaolei (Department of Clinical Laboratory Science,Guiyang Medical College,Guiyang 550004,Guizhou,China)

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DOI: 10.19367/j.cnki.1000-2707.2011.01.006

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