Mutation detection was performed using Reverse Dot Blot (RDB) procedure, as described by Lappin et al. In this study all 5 mutations were screened simultaneously using reverse dot blot (RDB) assay.ĭNA from the leukocytes of peripheral blood was prepared by the salting out protocol. RDB is more advantageous than other techniques since it does not necessitate the use of radiolabled probes as in Allele Specific Oligonucleotide assay and the use of positive controls to show the proper functioning of PCR reaction in each tube as in ARMS assays, since in ARMS technique, results are based on the absence of PCR products. This technique was evaluated as routine first-line analyses of the CFTR gene status. The reverse dot-blot (RDB) technique is one of the most widely used techniques to diagnose CF. Several techniques such as allele specific oligonucleotide (ASO) dot-blot, reverse dot-blot, amplification refractory mutation (ARMS), and an oligo-ligation assay, are available to detect the most common mutations. The knowledge of the mutation spectrum and its frequency in this population will improve the diagnosis and medical care of CF patients and could also be used as a basis for carrier screening and prenatal diagnosis programming in Iran. We selected five mutations, deltaF508, N1303K, G542X, R347H and W1282X based on previous reports in Iran and neighboring countries. In addition, the clinical presentations and laboratory findings were studied based on hospital and outpatient records. In the present study, 5 of the 27 CFTR exons of 30 unrelated northern Iranian CF patients were analyzed with the aim of detecting common CFTR mutations in this population. Intriguingly, p.G542X and p.N1303K are found on the same haplotype background as ΔF508, suggesting that they arose in the same population.įew previous reports of CFTR mutations in Iran have been published. Only four (p.G542X, p.N1303K, p.G551D and p.W1282X) have overall frequencies higher than 1%. These mutations vary greatly in their frequency and distribution, but most are very rare. In addition to the ΔF508 mutation, more than 1000 other mutations in the CFTR gene have been identified (CF Genetic Analysis Consortium. The p.F508del (ΔF508) allele of the gene is the most common mutation observed worldwide and is probably very old, dating to pre-Neolithic times. Other clinical features are variably associated with the disease.Īpproximately one in 2000 to 3000 newborns in populations of European ancestry are affected, and the average carrier frequency is about 1:25. Most of the patients also fail to produce digestive enzymes in the pancreas, resulting in pancreatic insufficiency. Excess mucus in the respiratory system of CF patients facilitates chronic bacterial infections, leading to respiratory failure, which is the major cause of mortality. The CFTR channel is critical for the normal function of epithelial cells in the lungs, pancreas, intestine, gall bladder, and sweat glands. The protein product of the gene is a chloride channel and a member of the ATP-binding cassette membrane transporter superfamily. ![]() Cystic fibrosis (CF) is an autosomal recessive disease caused by defect in the cystic fibrosis transmembrane conductance regulator (CFTR) gene.
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