Putative digenic forms.

Almost 40% of infertile men cases are classified as idiopathic when tested negative to the current diagnostic routine based on the screening of karyotype, Y chromosome microdeletions and CFTR mutations in men with azoospermia or oligozoospermia. Rare monogenic forms of infertility are not routinely evaluated. In this study we aim to investigate the unknown potential genetic causes in couples with pure male idiopathic infertility by applying variant prioritization to whole exome sequencing (WES) in a cohort of 99 idiopathic Italian patients. The ad-hoc manually curated gene library prioritizes genes already known to be associated with more common and rare syndromic and non-syndromic male infertility forms. Twelve monogenic cases (12.1%) were identified in the whole cohort of patients. Of these, three patients had variants related to mild androgen insensitivity syndrome, two in genes related to hypogonadotropic hypogonadism, and six in genes related to spermatogenic failure, while one patient is mutant in PKD1. These results suggest that NGS combined with our manually curated pipeline for variant prioritization and classification can uncover a considerable number of Mendelian causes of infertility even in a small cohort of patients.针对无精症（azoospermia）或少精症（oligozoospermia）男性患者，现行诊断流程通过染色体核型筛查、Y染色体微缺失检测及囊性纤维化跨膜传导调节因子（CFTR）基因突变检测进行评估，其中约40%的不育男性病例检测结果呈阴性，被归类为特发性不育。罕见单基因性不育尚未纳入常规临床评估流程。本研究纳入99名意大利特发性不育男性患者作为队列，通过全外显子组测序（whole exome sequencing, WES）开展变异优先级分析，旨在探究单纯男性特发性不育夫妇中未知的潜在遗传致病因素。本研究搭建的定制化手工整理注释基因库，可对已被证实与常见、罕见综合征性及非综合征性男性不育相关的基因进行优先级排序。在全部患者队列中，共检出12例单基因性不育病例（占比12.1%）。其中，3例患者携带与轻度雄激素不敏感综合征相关的变异；2例患者的变异涉及低促性腺激素性性腺功能减退症相关基因；6例患者的变异与生精障碍相关基因相关；另有1例患者存在PKD1基因突变。上述结果表明，即便针对小规模患者队列，将下一代测序（next-generation sequencing, NGS）与本研究的手工整理注释变异优先级分析及分类流程相结合，仍可检出大量不育症的孟德尔式致病原因。