title: Mutation characterization of CFTR gene in 206 Northern Irish CF families: thirty mutations, including two novel, account for approximately 94% of CF chromosomes.
abstract: A variety of mutation detection techniques, including restriction endonuclease digestion, allele specific oligonucleotides, and automated fluorescent sequencing, were used in the identification of 15 CFTR mutations representing 86.7% of CF chromosomes in 206 Northern Irish cystic fibrosis (CF) families. A systematic analysis of the 27 exons and intron/exon boundaries of the CFTR gene was performed using denaturing gradient gel electrophoresis (DGGE) in an attempt to characterise the 55 unknown CF mutations in 51 patients. Twenty different mutations were detected by DGGE on 30 chromosomes accounting for a further 7.3% of CF alleles. Fifteen of these mutations had not previously been found in Northern Ireland, and two are novel, M1I(G > T) and V562L. In total, 30 CFTR mutations account for 93.9% of the 412 Northern Irish CF chromosomes tested. The three major CF mutations in Northern Ireland are delta F508, G551D, and R117H with respective frequencies of 68.0%, 5.1%, and 4.1%. The efficacy of the DGGE technique was proven by the detection of 77 out of 77 control variants from all the CFTR exons. DGGE is a highly efficient and sensitive method for mutation screening especially in large genes where the mutation spectrum is known to be heterogeneous.
authors.affiliation: Queen's University of Belfast, Northern Ireland.
authors.forename: D J
authors.initials: DJ
authors.lastname: Hughes
authors.forename: A J
authors.initials: AJ
authors.lastname: Hill
authors.forename: M
authors.initials: M
authors.lastname: Macek
authors.forename: A O
authors.initials: AO
authors.lastname: Redmond
authors.forename: N C
authors.initials: NC
authors.lastname: Nevin
authors.forename: C A
authors.initials: CA
authors.lastname: Graham
mesh_terms: D000483:Alleles; D003550:Cystic Fibrosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004251:DNA Transposable Elements; D005091:Exons; D006801:Humans; D007438:Introns; D009154:Mutation; D009660:Northern Ireland; D009838:Oligodeoxyribonucleotides; D017354:Point Mutation; D011110:Polymorphism, Genetic; D015183:Restriction Mapping; D017384:Sequence Deletion; D013050:Spectrometry, Fluorescence
chemical_list: C505032:CFTR protein, human; D004251:DNA Transposable Elements; D009838:Oligodeoxyribonucleotides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator