一种新型通用引物多重PCR技术可特异性检测Y染色体序列标签位点

目的:建立通用引物多重PCR(UP-M-PCR)技术,检测Y染色体微缺失。方法:选取50例健康男性,针对Y染色体AZFa,AZFb,AZFc和AZFd区15个标签位点(STS),选择15对特异性引物和1对UP,将UP加在特异性引物对的5’端,按扩增片段大小组建4组稳定可靠的UP-M-PCR体系,分析其扩增效率和特异性。结果:UP引入M-PCR对检测体系无影响。UP-M-PCR可特异性扩增AZF区15个STS,且较M-PCR特异性更好,条带更清晰。结论:UP-M-PCR体系较M-PCR操作方便,扩增效率均衡,特异性高,是男性不育患者Y染色体微缺失筛查的新方法。

A Novel Universal Multiplex PCR Technology Could Specifically Detect Y Chromosome Sequence-Tagged-Sites

Objective：In order to improve the specificity and accuracy of Y chromosome microdeletion＇s assay, the study will explore a novel universal multiplex PCR instead of traditional multiplex PCR,which will be used for detection of Y chromosome microdeletions.Method：50 healthy men were selected for the study,for detection of 15 tagged sites （STS）of AZFa,AZFb,AZFc and AZFd district in Y chromosome,the study selected 1 5 pairs of spe-cific primers and one pair of universal primers （UP）.The UP was added to the 5＇end of each specific primer,and es-tablished 4 stable and reliable multiplex PCR reaction sets.Results：The data showed that UP-M-PCR method could specifically amplify the 1 5 STS,and we detected that UP-M-PCR could amplify the 1 5 STS more specifically and ac-curately as well as much clearer banes compared with the traditional M-PCR.Conclusion：The novel universal prim-er-multiplex-PCR （U-M-PCR）method can overcome the disadvantages of traditional multiplex PCR （M-PCR） which showed complex operation,poor specificity and low amplification efficiency.Thus U-M-PCR was a new anal-ysis method for routine screening of male infertility associated with Y chromosome microdeletions.