Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padua, Via Giustiniani, 2, 35128, Padova, Italy.
Familial Cancer Clinic, Veneto Institute of Oncology (IOV-IRCCS), Padua, Italy.
J Assist Reprod Genet. 2020 Apr;37(4):753-762. doi: 10.1007/s10815-020-01747-0. Epub 2020 Apr 3.
To develop and assess a novel custom next-generation sequencing (NGS) panel for male infertility genetic diagnosis.
A total of 241 subjects with diagnosis of idiopathic infertility ranging from azoospermia to normozoospermia were sequenced by a custom NGS panel including AR, FSHB, FSHR, KLHL10, NR5A1, NANOS1, SEPT12, SYCP3, TEX11 genes. Variants with minor allele frequency < 1% were confirmed by Sanger sequencing.
Nineteen missense variants were detected in 23 subjects with abnormal sperm count, whilst no variants were identified in normozoospermic men. Of identified variants, we prioritized variants classified as pathogenic and of uncertain significance (VUS) (63.1%, 12/19). No missense variants were found in males with normal seminal parameters (0/67). Therefore, the prevalence of variants was significantly higher in patients with spermatogenic impairment (16/174 vs 0/67, p = 0.007).
This study confirms the utility to apply NGS panel for infertility diagnosis in order to find new genetic variants potentially linked to male infertility with much higher accuracy than standard tests suggested by guidelines. Indeed, based on biological significance, prevalence in the general population and clinical data of patients, it is plausible that identified variants in this study might be linked to quantitative spermatogenic impairment, although further studies are needed.
开发和评估一种用于男性不育遗传诊断的新型定制下一代测序（NGS）面板。
对 241 例从无精子症到正常精子症的特发性不育症患者进行了一项定制 NGS 面板的测序，该面板包括 AR、FSHB、FSHR、KLHL10、NR5A1、NANOS1、SEPT12、SYCP3、TEX11 基因。通过 Sanger 测序证实了等位基因频率<1%的变异。
在 23 例精子计数异常的患者中检测到 19 种错义变异，而在正常精子症患者中未发现变异。在确定的变异中，我们优先考虑归类为致病性和意义不明的变异（VUS）（63.1%，12/19）。在精子参数正常的男性中未发现错义变异（0/67）。因此，在生精功能障碍患者中变异的发生率明显更高（16/174 与 0/67，p=0.007）。
本研究证实了在不孕症诊断中应用 NGS 面板的实用性，以便以比指南推荐的标准测试高得多的准确性找到与男性不育症相关的新遗传变异。事实上，根据生物学意义、在普通人群中的流行率和患者的临床数据，本研究中发现的变异很可能与数量性生精功能障碍有关，尽管还需要进一步的研究。
