应用靶向高通量测序技术检测不育男性的 Y 染色体结构变异。
Y chromosome structural variation in infertile men detected by targeted next-generation sequencing.
机构信息
Center for Reproductive Medicine, Center of Prenatal Diagnosis, First Hospital, Jilin University, 71th. Xin-min Street, Changchun, 130021, Jilin Province, People's Republic of China.
Peking Medriv Academy of Genetics and Reproduction, Peking, 102629, China.
出版信息
J Assist Reprod Genet. 2021 Apr;38(4):941-948. doi: 10.1007/s10815-020-02031-x. Epub 2021 Jan 16.
PURPOSE
To provide a validated method to identify copy number variation (CNV) in regions of the Y chromosome of infertile men by next-generation sequencing (NGS).
METHODS
Semen analysis was used to determine the quality of semen and diagnose infertility. Deletion of the azoospermia factor (AZF) region in the Y chromosome was detected by a routine sequence-tagged-site PCR (STS-PCR) method. We then used the NGS method to detect CNV in the AZF region, including deletions and duplications.
RESULTS
A total of 326 samples from male infertility patients, family members, and sperm donors were studied between January 2011 and May 2017. AZF microdeletions were detected in 120 patients by STS-PCR, and these results were consistent with the results from NGS. In addition, of the 160 patients and male family members who had no microdeletions detected by STS-PCR, 51 cases were found to exhibit Y chromosome structural variations by the NGS method (31.88%, 51/160). No microdeletions were found in 46 donors by STS-PCR, but the NGS method revealed 11 of these donors (23.91%, 11/46) carried structural variations, which were mainly in the AZFc region, including partial deletions and duplications.
CONCLUSION
The established NGS method can replace the conventional STS-PCR method to detect Y chromosome microdeletions. The NGS method can detect CNV, such as partial deletion or duplication, and provide details of the abnormal range and size of variations.
目的
提供一种通过下一代测序（NGS）鉴定不育男性 Y 染色体区域拷贝数变异（CNV）的验证方法。
方法
精液分析用于确定精液质量并诊断不育症。通过常规序列标记位点 PCR（STS-PCR）方法检测 Y 染色体无精子因子（AZF）区域的缺失。然后，我们使用 NGS 方法检测 AZF 区域的 CNV，包括缺失和重复。
结果
2011 年 1 月至 2017 年 5 月，共研究了 326 例男性不育症患者、家庭成员和精子供体的样本。STS-PCR 检测到 120 例患者存在 AZF 微缺失，结果与 NGS 一致。此外，在 160 例经 STS-PCR 检测无微缺失的患者和男性家庭成员中，51 例通过 NGS 方法发现 Y 染色体结构变异（31.88%，51/160）。STS-PCR 未在 46 名供体中发现微缺失，但 NGS 方法发现其中 11 名（23.91%，11/46）携带结构变异，主要位于 AZFc 区，包括部分缺失和重复。
结论
建立的 NGS 方法可以替代传统的 STS-PCR 方法来检测 Y 染色体微缺失。NGS 方法可以检测 CNV，如部分缺失或重复，并提供变异的异常范围和大小的详细信息。
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