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目的比较STS筛查方案与基因特异筛查方案在男性不育患者Y染色体微缺失检测中的效率。方法采用STS筛查方案与候选基因筛查方案对423例男性不育患者进行Y染色体微缺失检测。STS筛查方案依据EAA/EMQN推荐的方法进行,选择6个STS位点,分别为AZFa(SY84,SY86);AZFb(SY127,SY134);AZFc(SY254,SY255);基因特异筛查方案选择5个候选基因,分别为AZFa区(USP9Y、DBY),AZFb区(RBM1、SMCY),AZFc区(DAZ)。对实验中发现的SY84孤立缺失的病例进行验证,分析其引物序列区域的两个SNP(上游引物区域中SNP:5’+9insC和下游引物中的SNP:5’+15T>G(rs72609647))对检测结果的影响。结果两种筛查方案在423例患者均检测到30例相同的缺失,AZFc缺失21例;AZFb和AZFc共缺失为6例;AZFb缺失为1例;AZFa+b+c共缺失病例2例。STS位点筛查方案中有8例患者存在SY84位点的孤立缺失,进一步验证发现其是由于上游引物中的插入错配和下游引物序列中的SNP位点(rs72609647)变异导致其不能有效扩增导致。结论 STS筛查方案与基因特异筛查方案在Y染色体微缺失检测中,对于AZFb区和AZFc缺失检测中检测效率是一致的,但是在AZFa区缺失的检测中STS筛查方案存在假阳性。候选基因筛查方案较STS筛查方案更有利于临床推广应用。
Objective To compare the efficiency of STS screening and gene-specific screening in detection of Y chromosome microdeletions in male infertility patients. Methods 423 male infertility patients were tested for Y chromosome microdeletion by using STS screening program and candidate gene screening program. The STS screening protocol was based on the EAA / EMQN recommended method. Six STS sites were selected, which were AZFa (SY84, SY86), AZFb (SY127, SY134) and AZFc (SY254, SY255) The candidate genes were AZFa (USP9Y, DBY), AZFb (RBM1, SMCY) and AZFc (DAZ). Two SNPs in the primer sequence region (SNP: 5 ’+ 9insC in the upstream primer region and SNP: 5’ + 15T> G (rs72609647) in the downstream primer) were validated in the case of SY84 isolated deletion found in the experiment. Impact on test results. Results There were 30 cases of the same deletion in 423 cases, 21 cases of AZFc deletion, 6 cases of AZFb and AZFc deletion, 1 case of AZFb deletion and 2 cases of AZFa + b + c deletion. In the STS locus screening program, 8 patients had isolated deletion of SY84 locus. Further validation revealed that they were not effective due to insertion mismatch in the upstream primer and mutation of the SNP locus (rs72609647) in the downstream primer sequence Increased lead. Conclusions STS screening and gene-specific screening programs are consistent in detection of AZFb deletion and AZFc deletion in Y chromosome microdeletions, but there is a false positive for STS screening in AZFa deletion testing. Candidate gene screening program STS screening program is more conducive to the clinical application.