NGS技术在非梗阻性无精子症和少精子症患者Y染色体AZF区检测中的应用  

The application of NGS technology for detecting AZF region on the Y chromosome in non-obstructive patients with azoospermia and oligozoospermia

作　　者：高玲霞 王玲 蔡杨 马旭辉 李建波 王珺 Gao Lingxia;Wang Ling;Cai Yang;Ma Xuhui;Li Jianbo;Wang Jun(Department of Reproductive Medicine Center,Tangdu Hospital of The Fourth Military Medical University,Xi'an 710038)

机构地区：[1]空军军医大学唐都医院妇产科生殖医学中心

出　　处：《中国男科学杂志》2019年第1期3-8,共6页Chinese Journal of Andrology

基　　金：国家自然科学基金(31640056);陕西省自然科学面上项目(2016JM8052)

摘　　要：目的探讨NGS技术在非梗阻性无精子症及少精子症患者AZF区域基因重排检测中的应用可能,同时初步探讨AZF区拷贝数变化(CNV)与男性生精障碍的关系。方法选取来自唐都医院生殖医学中心就诊的53例非梗阻性无精子症和少精子症为实验组,219例精子数目正常男性为对照组,采用Ion Proton技术平台,通过基于选定区域捕获及NGS技术进行AZF区域基因重排分析,所有样本同时行AZF序列标签位点的多重PCR检测(STS-PCR)。结果实验组中STS-PCR检出率为11.32%(6/53),NGS技术检出率为52.83%(28/53).对照组中STS-PCR检出率为0%(0/219),NGS技术检出率为25.57%(56/219)。统计分析结果表明gr/gr缺失在实验组和对照组之间具有统计学差异(P=0.022),AZFc区完全缺失在实验组和对照组之间具有统计学差异(P=0.007)。其他CNV在实验组和对照组之间没有统计学差异。结论基于选定区域捕获结合NGS技术具有更高的灵敏度和分辨率,是进行AZF区基因重排分析检测的一种精确可靠的技术;gr/gr缺失和AZFc区完全缺失与精子生成相关。Objective To investigate the possibility of applyingthe selected area captureand NGS tothe detection of azoospermia factor (AZF) microdeletion on the Y chromosome in infertile men with azoospermia or severe oligozoospermia. Meanwhile, we also preliminarily discuss the relationship between CNV of AZF area and spermatogenesis impairment Methods We collect 53 non-obstructive patients with azoospermia and oligozoospermia as the experimental group, and 219 cases with normal semen as control group from reproductive medicine center, Tangdu hospital, analysing gene rearrangement of AZF area via the selected area capture andNGSwith Ion Proton platform. Meanwhile, all samples also are analysed by the AZF sequence label locus PCR. Results The detection rate of CNV of the experimental group is 11.32% by using STS-PCR, and NGS is 52.83%. The detection rate of CNV of the control group is 0% by using STS-PCR, and NGS is 25.57%. The gr/gr deletion and complete deletion of AZFc are statistically significantbetween the two groups (P <0.05), other CNVs in this study are not statistically significant between the two groups. Conclusion NGS is a high sensitivity and specificity method, which is used to detectgene rearrangement of AZF area. Gr/gr deletion andcomplete deletion of AZFcare associated with spermatogenesis.

关 键 词：无精子症因子 少精子症 下一代基因测序技术 

分 类 号：R679[医药卫生—外科学]