SNaPshot assay for the detection of the most common CFTR mutations in infertile men.
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.
Macedonian Academy of Sciences and Arts, Research Center for Genetic Engineering and Biotechnology 'Georgi D. Efremov', Skopje, Republic of Macedonia.
Predrag
P
Noveski
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.
Svetlana
S
Madjunkova
Macedonian Academy of Sciences and Arts, Research Center for Genetic Engineering and Biotechnology 'Georgi D. Efremov', Skopje, Republic of Macedonia.
Marija
M
Mircevska
Faculty of Medicine, Clinic of Endocrinology and Metabolic Disorders, Skopje, Republic of Macedonia.
Toso
T
Plaseski
Clinical Hospital 'Acibadem Sistina', Skopje, Republic of Macedonia.
Vanja
V
Filipovski
Macedonian Academy of Sciences and Arts, Research Center for Genetic Engineering and Biotechnology 'Georgi D. Efremov', Skopje, Republic of Macedonia.
Dijana
D
Plaseska-Karanfilska
D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004252:DNA Mutational Analysis; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006801:Humans; D007248:Infertility, Male / Q000235:genetics*; D008297:Male
C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator