两种试剂盒DYS531分型差异分析

Study on the different typing results between two kits for DYS531 Locus

目的 探究 AGCU Y SUPP PLUS 和 GoldeneyeY ADD 两个试剂盒在 DYS531 基因座出现分型差异的原因。 方法 用 AGCU Y SUPP PLUS、GoldeneyeY ADD 试剂盒检测 222 份随机样本及 007、9948 标准品,随机抽取两试剂盒中 DYS531 基因座分型存在差异的 7 份样本及分型一致的 1、211、212 号样本,对 DYS531 基因座进行 Sanger 测序。 结果 两种试剂盒的电泳结果显示,1、211、212 号样本 DYS531 基因座分型相同,剩余 219 份样本存在 5 碱基差异;测序结果表明,随机抽取分型有差异的 7 个样本在上游侧翼序列 rs58571108 位置上均存在GCATG 5 碱基插入,1、211、212 号样本不存在该插入。结论 AGCU Y SUPP PLUS 和 GoldeneyeY ADD 两个试剂盒分型差异是由上游侧翼序列的 rs58571108 导致,AGCU Y SUPP PLUS 在引物设计时避开了该多态性位点,分型准确可靠。

Objective To explore the reasons for the genotyping differences between AGCU Y SUPP PLUS and GoldeneyeTM Y ADD kits for the DYS531 locus. Methods 222 random samples and two standards(007 and 9948) were typed by AGCU Y SUPP PLUS kit and GoldeneyeTM Y ADD kit respectively. Sanger sequencing was performed on the DYS531locus for 7 randomly selected samples with different STR results and samples No.1, No.211 and No.212 with identical types.Results The capillary electrophoresis results of the two domestic kits showed that samples No.1, No.211 and No.212 had identical typing results for DYS531, while the remaining 219 samples had 5 base pairs differences. The sequencing results demonstrated that 7 randomly selected samples with different STR results had GCATG insert(rs58571108) at the upstream flanking sequence, which did not exist for samples No.1, No.211 or No.212. Conclusion The typing difference between AGCU Y SUPP PLUS kit and GoldeneyeTM Y ADD kit was caused by rs58571108 on the upstream flanking sequence. AGCU Y SUPP PLUS kit avoided this polymorphic locus in the primer design, which results in accurate and reliable typing results.