title: Analysis of the complete coding region of the CFTR gene in a cohort of CF patients from north-eastern Italy: identification of 90% of the mutations.
abstract: A complete coding-region analysis on 225 cystic fibrosis (CF) chromosomes from a cohort that includes all the affected subjects born in two North-Eastern Italian regions over eight years was performed. In a previous study, we identified mutations on 166/225 (73.8%) CF chromosomes after screening for 62 mutations. To characterise the remaining 59 CF chromosomes, we carried out automated direct DNA sequencing (exons 9 and 13), RNA single-strand conformation polymorphism (exons 1-8 and 10-12) and denaturing gradient gel electrophoresis (exons 14a-24) of the 27 exons and flanking regions of the CF transmembrane conductance regulator gene. We identified 22 mutations, four of which are novel, viz. 711 + 5G-->A, R709X, 3132delTG and 2790-2A-->G, and we characterised 90.2% (203/225) of the CF chromosomes. Taking advantage of the homogeneity of the sample, an evaluation of the most important clinical parameters, assessed at the age of 12 years, is presented. We confirm some previously reported genotype-phenotype correlations and we report a new nonsense mutation (R709X) associated with a pancreatic sufficient phenotype.
authors.affiliation: Centro Regionale Veneto Fibrosi Cistica, Ospedale Civile Maggiore, Verona, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Bonizzato
authors.forename: L
authors.initials: L
authors.lastname: Bisceglia
authors.forename: C
authors.initials: C
authors.lastname: Marigo
authors.forename: E
authors.initials: E
authors.lastname: Nicolis
authors.forename: C
authors.initials: C
authors.lastname: Bombieri
authors.forename: C
authors.initials: C
authors.lastname: Castellani
authors.forename: G
authors.initials: G
authors.lastname: Borgo
authors.forename: L
authors.initials: L
authors.lastname: Zelante
authors.forename: G
authors.initials: G
authors.lastname: Mastella
authors.forename: G
authors.initials: G
authors.lastname: Cabrini
mesh_terms: D000293:Adolescent; D000328:Adult; D001483:Base Sequence; D002648:Child; D003062:Codon; D015331:Cohort Studies; D003550:Cystic Fibrosis / Q000453:epidemiology / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004252:DNA Mutational Analysis; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D006801:Humans; D007558:Italy / Q000453:epidemiology; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D016133:Polymerase Chain Reaction
chemical_list: C505032:CFTR protein, human; D003062:Codon; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator