title: [Detection of mutations in the CFTR gene using the PCR-dependent preferential homoduplex formation assay (PCR-PHFA) system].
abstract: A simple and efficient method for detecting DNA alterations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene has been developed using polymerase chain reaction dependent preferential homoduplex formation assay (PCR-PHFA). The technique is based on strand competition between a double labeled amplicon and an unlabeled amplicon during hybridization in a precisely controlled temperature gradient. By using this method the presence of single base substitutions in a PCR amplicon can be detected. In this report, experimental evaluation of the PCR-PHFA system in DNA analysis of exons of the CFTR gene with the highest frequency of most common disease-causing mutations is presented. The method was capable of detecting all mutations tested within the amplified regions of genomic DNA. PCR-PHFA can be used for mutation screening of CFTR and potentially other genes with a large number of reported mutations.
authors.affiliation: Institute for Biotechnology Research, Wakunaga Pharmaceutical Co, Ltd.
authors.forename: T
authors.initials: T
authors.lastname: Oka
authors.forename: H
authors.initials: H
authors.lastname: Nakano
authors.forename: A
authors.initials: A
authors.lastname: Yamane
mesh_terms: D003550:Cystic Fibrosis / Q000175:diagnosis*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D006801:Humans; D008722:Methods; D009154:Mutation; D016133:Polymerase Chain Reaction
chemical_list: C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator