title: Screening for cystic fibrosis gene mutations by multiplex DNA amplification.
abstract: We have developed a simple rapid DNA screening test that allows us simultaneously to analyze seven CF mutations (delta F508, R347P, S549N, G551D, R553X, R334W, 444delA) that together account for about 60% of all CF mutations in the Italian population. It consists of three steps: multiplex polymerase chain reaction (PCR) amplification of exons 4, 7, 10 and 11; restriction endonuclease digestion of the PCR products; and vertical polyacrylamide gel electrophoresis analysis. We have used our multiplex assay for analyzing 15 CF chromosomes (non delta F508) and have found 3 cases of the R553X mutation; the latter have been confirmed by amplification and digestion of exon 11.
authors.affiliation: Department of Pediatrics, University of Padua, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Picci
authors.forename: F
authors.initials: F
authors.lastname: Anglani
authors.forename: M
authors.initials: M
authors.lastname: Scarpa
authors.forename: F
authors.initials: F
authors.lastname: Zacchello
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003550:Cystic Fibrosis / Q000235:genetics*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005091:Exons; D006580:Genetic Carrier Screening; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D012016:Reference Values; D015183:Restriction Mapping
chemical_list: D009838:Oligodeoxyribonucleotides; D004247:DNA