无精子症患者基因检测分析

目的:利用全外显子测序技术,筛选无精子症患者相关基因,丰富男性不育基因库。方法:抽取20例无精子症患者外周血,提取DNA,使用杂交捕获方法构建DNA文库,采用高通量测序技术检测人类全外显子组中20099个基因的外显子区及旁侧内含子区(20bp),将测序数据与人类基因组hg19参考序列进行比对,筛选变异基因,变异位点进行Sanger测序验证。结果:共计筛选出26个基因43个变异位点,排除15个常染色体隐性遗传的单个变异位点及13个与精子运动相关的变异位点,剩余11个基因的15个变异位点可能与精子发生障碍相关,其中包括FAM71B、STARD9、CLTCL1、PCBP3、S100PBP等5个在睾丸组织高表达基因,SYCE3、EFCAB6、DDX4、KDM5D、RGS22、MTL5等6个基因可能与精子发生障碍相关。结论:通过研究筛选到可能影响男性不育的基因,为男性不育的基因诊断研究提供参考。

Analysis of genetic testing of patients with azoospermia

Objective:To screen genes related to azoospermia by exome sequencing technology,and to enrich male infertility gene pool.Methods:Peripheral blood of 20 patients with azoospermia was collected,and DNA was extracted from their peripheral blood.DNA libraries were constructed by hybrid capture methods,and high-throughput sequencing technology was used to detect the exon regions and flanking contents of 20099 genes in the human exome subregion(20bp),the sequencing data was compared with the human genome hg19 reference sequence,and the mutant genes were screened.The mutation sites were verified by Sanger sequencing.Results:A total of 43 mutation sites of 26 genes were screened out,and 15 autosomal recessive single mutation sites and 13 mutation sites related to sperm motility were excluded.The remaining 15 mutation sites of 11 genes may be related to the disorders of spermatogenesis,which included 5 genes,such as FAM71B,STARD9,CLTCL1,PCBP3,S100PBP,were high expression in testicular tissue.And 6 genes including SYCE3,EFCAB6,DDX4,KDM5D,RGS22,and MTL5 could be related to spermatogenesis disorders.Conclusion:The genes that may affect male infertility have be screened in this research,which can provide evidence for the genetic diagnosis of male infertility.