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.

中文翻译：

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靶向二代测序检测不育男性Y染色体结构变异

目的

提供一种通过下一代测序 (NGS) 识别不育男性 Y 染色体区域拷贝数变异 (CNV) 的有效方法。

方法

精液分析用于确定精液质量和诊断不孕症。通过常规序列标记位点 PCR (STS-PCR) 方法检测 Y 染色体中无精子因子 (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 份来自男性不育患者、家庭成员和精子捐献者的样本。通过 STS-PCR 在 120 名患者中检测到 AZF 微缺失，这些结果与 NGS 的结果一致。此外，STS-PCR检测未发现微缺失的160例患者及男性家属中，NGS方法发现Y染色体结构变异51例（31.88%，51/160）。STS-PCR在46名供体中未发现微缺失，但NGS方法显示这些供体中有11名（23.91％，11/46）携带结构变异，主要在AZFc区域，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方法检测Y染色体微缺失。NGS 方法可以检测 CNV，例如部分缺失或重复，并提供异常范围和变异大小的详细信息。