ABCMdb

PMID: 21976147

Safinejad K, Darbouy M, Kalantar SM, Zeinali S, Mirfakhraie R, Yadegar L, Houshmand M
The prevalence of common CFTR mutations in Iranian infertile men with non-CAVD obstructive azoospermia by using ARMS PCR techniques.
J Assist Reprod Genet. 2011 Oct 6., [PubMed]

Sentences

No. Mutations Sentence Comment

0 ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* GENETICS The prevalence of common CFTR mutations in Iranian infertile men with non-CAVD obstructive azoospermia by using ARMS PCR techniques Kyumars Safinejad & Mojtaba Darbouy & Sayed Mahdi Kalantar & Sirus Zeinali & Reza Mirfakhraie & Leila Yadegar & Masoud Houshmand Received: 9 May 2011 /Accepted: 24 August 2011 /Published online: 6 October 2011 # Springer Science+Business Media, LLC 2011 Abstract Purpose To evaluate five common cystic fibrosis transmembrane conductance regulator (CFTR) mutations (ΔF508, G542X, R117H, W1282X and N1303K) in the Iranian infertile men with noncongenital absence of vas deferens (CAVD) obstructive azoospermia. Login to comment 3 ABCC7 p.Gly542* Results The common CFTR mutations were found positive in 5/53)9.43%(for ΔF508 and 4/53)7.55%(for G542X mutation of all patients tested. Login to comment 34 ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* The aim of this study was to evaluate five common CF mutations (ΔF508, G542X, R117H,W1282X, N1303K)by use of the multiplex and single ARMS system among Iranian men with non-CAVD obstructive azoospermia (including those with idiopathic epididymal or ejaculatory duct obstruction) as the first descriptive study. Login to comment 38 ABCC7 p.Arg117His ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Gly542* The diagnosis of non-CAVD obstructive azoospermia is based on the following examinations: normal semen volume; normal testicular size; presence of the vas deferens by clinical examination; normal levels of serum follicle-stimulating hormone (FSH);azoospermia; absence or low levels of fructose and presence of spermatozoa in sample extracted by percutaneous sperm aspiration(PESA).No other symptoms of CF such as chronic lung inflammation/infection, pancreatic Table 1 Allelic and Genotypic Frequencies in Iranian infertile men with non-CAVD obstructive azoospermia Mutation No. of chromosomes carry CF allele %(Allelic frequencies) Genotype No. of patients %(Genotypic frequencies) ΔF508 5/106 4.7 ΔF508/+ 5 9.43 G542X 4/106 3.77 G542X/+ 4 7.55 R117H 0/106 0 R117H/+ 0 0 W1282X 0/106 0 W1282X/+ 0 0 N1303K 0/106 0 N1303K/+ 0 0 Normal 97/106 91.5 +/+ 44 83 Total 106/106 100.00 Total 53 100.00 insufficiency and intestinal obstruction have been reported in clinical file of our patients. Login to comment 40 ABCC7 p.Arg117His ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Gly542* All DNA samples were analyzed, using the primer sequence and single and multiplex ARMS-PCR technique as described by Ferrie et al. [21], for the following mutations: ΔF508, N1303K, G542X,W1282X,R117H mutations.W1282X and R117H mutations were analyzed by single ARMS-PCR technique and ΔF508, N1303K and G542X mutations were analyzed simultaneously by multiplex ARMS-PCR technique. Login to comment 41 ABCC7 p.Arg117His ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ARMS PCR program for ΔF508, N1303K, G542X,R117H began with a 5 min incubation at 94°C,andProceeded with 28 cycles, each containing 15 s of denaturation at 94°C,30 s of annealing at appropriate temperature and 30 s of extension at 72°C;with a 10 min incubation at 72°C completing the amplification. Login to comment 42 ABCC7 p.Trp1282* ARMS PCR program for W1282X mutation began with a 1 min incubation at 95°C, and Proceeded with 32 cycles, each with 1 min of denaturation at 95°C, 30 s of annealing at appropriate temperature and 45 s of extension at 72°C; with a 5 min incubation at 72°C completing the amplification.PCR conditions for amplification of above DNA samples stood as described earlier [21]. Login to comment 43 ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Results Heterozygote frequency for ΔF508 mutation has been 5/53 (%9.43) and for G542X mutation, it has been 4/53(%7.55) in all patients tested where as other common mutations (R117H, W1282X, N1303K) were not detected in our samples. Login to comment 45 ABCC7 p.Gly542* In other words, frequency for ΔF508 mutation was 5/106 (%4.7) and for G542X mutation it proved 4/106(3.77) in all chromosomes tested. Login to comment 52 ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Among 53 patients with non-CAVD obstructive azoospermia, five were heterozygotes for ΔF508 mutation (9.43%), and four patients carried G542X mutation (7.55%) whereas other mutations (N1303K, W1282X andR117H) were not detected in our samples. Login to comment