Genotyping Efficiency of 2 Primer Sets and an Unlabeled Oligonucleotide Probe for the p.Phe508del in Exon 10 of the CFTR Gene as Determined with High-Resolution Melting Analysis

摘要

To the Editor:Cystic fibrosis (OMIM 219700) is one of the most common autosomal recessive disorders in Caucasians. It is caused by mutations in the CFTR 1 [cystic fibrosis transmembrane conductase regulator (ATP-binding cassette sub-family C, member 7)] gene, located on chromosome 7q31.2. The frequency of the most common mutation, p.Phe508del (F508del, c.1521_1523delCTT), varies markedly among ethnic groups, but is always tested in both patient testing and carrier screening (1, 2).At our institution, we screen for CFTR mutations with high-resolution melting analysis (LightScanner?; Idaho Technology). The method consists of PCR amplification of the fragment in question in the presence of a fluorescent dye (LCGreen Plus) that intercalates between bases in double-stranded DNA. The amplification products are melted after the PCR to reveal unique melting profiles according to their sequence compositions (3).A successful experiment depends on primer design (gene scanning) and probe design (genotyping), as well as on …