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[Detection of mutations in the CFTR gene using the PCR-dependent preferential homoduplex formation assay (PCR-PHFA) system].

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.