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

• 1:贵阳医学院检验系

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

Abstract:

Keywords:

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

DOI: 10.19367/j.cnki.1000-2707.2011.01.006

参考文献(References)：

• [1]LO YM,Corbetta N,Chamberlain PF,et al.PresenceoffetDNA in maternal plasma and serum[J].lancet,1997(350):485-487.
• [2]陈振斌,闫梅,雷箴,等.利用孕妇血浆DNA检测胎儿性别的研究[J].遗传,2004(1):18-22.
• [3]鹿亚超,朱斌,姜海燕,等.孕妇血浆Amelogenin基因6bp差异检测对产前胎儿性别的鉴定价值[J].苏州大学学报医学版,2005(3):416-418.
• [4]刘福民,冯霞,王秀英,等.荧光定量PCR检测孕妇血浆中胎儿DNA的研究[J].中华围产医学杂志,2005(6):380-382.
• [5]Hromadnikova I,Houbova B,Hridelova D,et al.Repli-cate realtime PCR testing of DNA in matemal plasma in-crease the sentitivity of non-invasive fetal sex determina-tion[J].Prenat Diagn,2003(23):235-238.
• [6]吕德坚,李建金,刘秋玲.中国汉族群体DXS101位点的多态性[J].中山医科大学学报,2001(22):1222-1241.
• [7]Lo YM,Lau TK,Poon PM,et al.Quantitative analysis offetal DNA in meternal plasma and serum:implications fornoninvasive prenatal diagnosis[J].Am J Hum Genent,1998(4):768-775.
• [8]易萍,李力,颜耀华,等.应用母亲血浆常规PCR技术进行胎儿性别鉴定[J].中国妇幼保健,2006(21):836-837.

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