title: Denaturing high-performance liquid chromatography quickly and reliably detects cardiac ion channel mutations in long QT syndrome.
abstract: Multiple mutations in several ion channel genes (KCNQ1, KCNH2, SCN5A, KCNE1, KCNE2, and KCNJ2) have been shown to cause autosomal dominant long QT syndrome (LQTS), a familial cardiac disorder that causes syncope, seizures, and sudden death. Due to their multiple loci and considerable size, mutation detection in these genes represents a challenge that is only partially met by the conventional screening method of single-stranded conformational polymorphism (SSCP). The recently introduced denaturing high-performance liquid chromatography (dHPLC) offers a promising new method for a fast and sensitive analysis of PCR-amplified DNA fragments. To test the applicability of dHPLC in the molecular diagnosis of LQTS, we first assessed a cohort of 192 patients from our International LQTS Registry for 14 previously identified mutations (including 10 different missense mutations, 1-bp, 2-bp, 3-bp, and 9-bp deletion mutations), and 2 polymorphisms in the LQTS potassium and sodium channel genes. Applying empirically determined exon-specific melting profiles, all mutations (including four previously undetectable by SSCP) were readily identified by dHPLC. We conclude that the dHPLC technology is a highly sensitive and efficient method for the molecular analysis of LQTS, and the same PCR amplicons developed for SSCP testing can be directly used for dHPLC assay.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, NY 14642, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Ning
authors.forename: Arthur
authors.initials: A
authors.lastname: Moss
authors.forename: Woject
authors.initials: W
authors.lastname: Zareba
authors.forename: Jennifer
authors.initials: J
authors.lastname: Robinson
authors.forename: Spencer
authors.initials: S
authors.lastname: Rosero
authors.forename: Dan
authors.initials: D
authors.lastname: Ryan
authors.forename: Ming
authors.initials: M
authors.lastname: Qi
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005820:Genetic Testing / Q000379:methods; D006321:Heart / Q000502:physiology; D006579:Heterozygote; D006801:Humans; D008133:Long QT Syndrome / Q000235:genetics*; D009154:Mutation; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D015221:Potassium Channels / Q000235:genetics*; D015203:Reproducibility of Results; D015222:Sodium Channels / Q000235:genetics*
chemical_list: D015221:Potassium Channels; D015222:Sodium Channels