title: Sensitivity of the denaturing gradient gel electrophoresis technique in detection of known mutations and novel Asian mutations in the CFTR gene.
abstract: More than 500 mutations have been identified in the CFTR gene, making it an excellent system for testing mutation scanning techniques. To assess the sensitivity of denaturing gradient gel electrophoresis (DGGE), we collected a representative group of 202 CFTR mutations. All mutations analyzed were detected by scanning methods other than the DGGE approach evaluated in this study. DGGE analysis was performed on 24 of the 27 exons and their flanking splice site sequences. After optimization, 201 of the 202 control samples produced an altered migration pattern in the region in which an alteration occurred. The remaining sample was sequenced and found not to have the reported mutation. The ability of DGGE to identify novel mutations was evaluated in three Asian CF patients with four unknown CF alleles. Three novel Asian mutations were detected-K166E, L568X, and 3121-2 A-->G (in homozygosity)-accounting for all CF alleles. These results indicate that an optimized DGGE scanning strategy is highly sensitive and specific and can detect 100% of mutations.
authors.affiliation: Center for Medical Genetics, CMSC 1004, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-3914, USA.
authors.forename: M
authors.initials: M
authors.lastname: Macek
authors.forename: B
authors.initials: B
authors.lastname: Mercier
authors.forename: A
authors.initials: A
authors.lastname: Macková
authors.forename: P W
authors.initials: PW
authors.lastname: Miller
authors.forename: A
authors.initials: A
authors.lastname: Hamosh
authors.forename: C
authors.initials: C
authors.lastname: Férec
authors.forename: G R
authors.initials: GR
authors.lastname: Cutting
mesh_terms: D000483:Alleles; D044466:Asian People / Q000235:genetics*; D003550:Cystic Fibrosis / Q000208:ethnology / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005091:Exons; D005260:Female; D006801:Humans; D008297:Male; D008957:Models, Genetic; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D012680:Sensitivity and Specificity
chemical_list: C505032:CFTR protein, human; D017931:DNA Primers; D019005:Cystic Fibrosis Transmembrane Conductance Regulator