title: Fluorescence-based, multiplex allele-specific PCR (MASPCR) detection of the delta F508 deletion in the cystic fibrosis transmembrane conductance regulator (CFTR) gene.
abstract: Cystic fibrosis (CF) is a common genetic disorder in Caucasians, and in some populations 70% of cases are associated with a 3 base pair (bp) deletion (delta F508) in the CFTR gene. We have implemented a fluorescence-based, multiplex allele-specific polymerase chain reaction (MASPCR) assay for deletion of the delta F508 mutation. Different allele-specific fluorescently-tagged primers are used in the PCR reaction to distinguish between normal and delta F508 alleles. Fluorescent PCR products are then visualized in a single lane on an agarose gel following electrophoresis combined with real-time multicolour fluorescence detection. The approach simplifies diagnosis of the most common mutation in the CFTR gene, and holds promise for a multiplex allele-specific, fluorescence-tagged gene amplification strategy for detection of additional CF mutations which may result in more cost-effective testing without increasing the risk of missed or erroneous diagnoses.
authors.affiliation: Molecular Biology Diagnostics Unit, Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine 19104.
authors.forename: P
authors.initials: P
authors.lastname: Fortina
authors.forename: R
authors.initials: R
authors.lastname: Conant
authors.forename: T
authors.initials: T
authors.lastname: Parrella
authors.forename: E
authors.initials: E
authors.lastname: Rappaport
authors.forename: T
authors.initials: T
authors.lastname: Scanlin
authors.forename: E
authors.initials: E
authors.lastname: Schwartz
authors.forename: J M
authors.initials: JM
authors.lastname: Robertson
authors.forename: S
authors.initials: S
authors.lastname: Surrey
mesh_terms: D000483:Alleles; D001483:Base Sequence; D003550:Cystic Fibrosis / Q000175:diagnosis*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005453:Fluorescence; D005809:Genes, Regulator / Q000235:genetics; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: C505032:CFTR protein, human; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator