Noveski Predrag, Madjunkova Svetlana, Mircevska Marija, Plaseski Toso, Filipovski Vanja, Plaseska-Karanfilska Dijana
Macedonian Academy of Sciences and Arts, Research Center for Genetic Engineering and Biotechnology 'Georgi D. Efremov', Skopje, Republic of Macedonia.
Macedonian Academy of Sciences and Arts, Research Center for Genetic Engineering and Biotechnology 'Georgi D. Efremov', Skopje, Republic of Macedonia; Division of Clinical Pharmacology and Toxicology, The Hospital for Sick Children, Toronto, Ontario, Canada.
PLoS One. 2014 Nov 11;9(11):e112498. doi: 10.1371/journal.pone.0112498. eCollection 2014.
Congenital bilateral absence of vas deferens (CBAVD) is the most common CFTR-related disorder (CFTR-RD) that explains about 1-2% of the male infertility cases. Controversial data have been published regarding the involvement of CFTR mutations in infertile men with non-obstructive azoospermia and oligozoospermia. Here, we describe single base extension (SNaPshot) assay for detection of 11 common CFTR mutations: F508del, G542X, N1303K, 621+1G->T, G551D, R553X, R1162X, W1282X, R117H, 2184insA and 1717-1G->A and IVS8polyT variants. The assay was validated on 50 previously genotyped samples and was used to screen a total of 369 infertile men with different impairment of spermatogenesis and 136 fertile controls. Our results show that double heterozygosity of cystic fibrosis (CF) and CFTR-related disorder (CFTR-RD) mutations are found in a high percentage (22.7%) of infertile men with obstructive azoospermia, but not in other studied groups of infertile men. The SNaPshot assay described here is an inexpensive, fast and robust method for primary screening of the most common CFTR mutations both in patients with classical CF and CFTR-RD. It can contribute to better understanding of the role of CFTR mutations in impaired spermatogenesis, ultimately leading to improved management of infertile men.
先天性双侧输精管缺如（CBAVD）是最常见的与囊性纤维化跨膜传导调节因子（CFTR）相关的疾病（CFTR-RD），约占男性不育病例的1%-2%。关于CFTR突变在非梗阻性无精子症和少精子症不育男性中的作用，已有争议性数据发表。在此，我们描述了一种用于检测11种常见CFTR突变的单碱基延伸（SNaPshot）分析方法：F508del、G542X、N1303K、621+1G→T、G551D、R553X、R1162X、W1282X、R117H、2184insA、1717-1G→A以及IVS8polyT变异体。该分析方法在50个先前已进行基因分型的样本上得到验证，并用于筛查总共369例患有不同程度精子发生障碍的不育男性以及136例生育力正常的对照者。我们的结果显示，在梗阻性无精子症不育男性中，发现囊性纤维化（CF）和CFTR相关疾病（CFTR-RD）突变的双杂合子比例很高（22.7%），但在其他所研究的不育男性群体中未发现。本文所述的SNaPshot分析方法是一种廉价、快速且可靠的方法，可用于对经典CF和CFTR-RD患者中最常见的CFTR突变进行初步筛查。它有助于更好地理解CFTR突变在精子发生障碍中的作用，最终改善对不育男性的管理。
