AMELY无效扩增个体Yp11.2区域STS位点检测

目的通过Y-STR复合扩增技术检测AMELY缺失的男性个体Y染色体的完整性,并对数据库中的STS位点进行筛选并设计引物,检测Y染色体AMEL区域的STS位点缺失情况,为进一步研究中国人口中AMELY的缺失提供数据和理论依据。方法应用Goldeneye 20A PCR扩增试剂盒、华夏TM白金PCR扩增试剂、Y filer plus试剂盒和Goldeneye 27YB试剂盒进行PCR扩增、通过ABI 3500遗传分析仪进行毛细管电泳以及STR分型。通过UniSTS database和UCSC对Y染色体p11.2区域的多个STS位点进行筛选,部分STS位点自行设计引物,梯度PCR和常规PCR扩增结合琼脂糖凝胶电泳进行STS位点的分析。结果 Goldeneye 20A和华夏TM白金两种试剂盒分型结果均显示两父子AMELY等位基因无效扩增。Y filer plus和27YB试剂盒分型结果显示两父子均在DYS570和DYS576位点无效扩增。STS位点扩增显示两父子的Y染色体缺失的STS位点有BV703904、BV703923、SY2137、SY716、DYS256、DYS267、SY2232、SY2233、DYS261和DYS260。结论 AMELY无效扩增的父子Yp11.2缺失片段包含了BV703904-DYS260之间的区域,长度为2.63~2.74 Mb。

Detection mapping of Y chromosome STS loci in AMELY dropout individuals

Objective Y-STR multiplex amplification technology is used to detect the integrity of the Y chromosome of male individuals with AMELY deletions, and the STS loci in database are screened and primers are designed to, detect the STSs in the AMEL delated region, so as to provide data and theoretical basis for further research on the deletion of AMELY in Chinese population. Methods Goldeneye 20 A PCR amplification kit, HuaxiaTM Platinum kit, Y filer plus kit and Goldeneye 27 YB kit for PCR amplification and ABI 3500 Gene Analyzer were used for capillary electrophoresis and STR genotyping. multiple STS loci in the Yp11.2 region were screened by UniSTS database and UCSC, some STSs designed their own primers., Gradient PCR and conventional PCR amplification combined with agarose gel electrophoresis were used for STSs analysis. Results The STR genotyping of the Goldeneye 20 A and HuaxiaTM Platinum kits both showed that the AMELY of the father and son were dropout. The genotyping results of Y filer plus and 27 YB kits showed that both father and son were negatively amplified at DYS570 and DYS576. Amplification of STSs showed that their Y chromosomes deletion STSs were BV703904, BV703923, SY2137, SY716, DYS256, DYS267, SY2232, SY2233, DYS261 and DYS260. Conclusions The Yp11.2 deletion region between father and son amplified by AMELY contains the region between BV703904-DYS260, which is 2.63~2.74 Mb in length.