title: Denaturing high-performance liquid chromatography screening of the long QT syndrome-related cardiac sodium and potassium channel genes and identification of novel mutations and single nucleotide polymorphisms.
abstract: Mutations in cardiac potassium and sodium channel genes are responsible for several hereditary cardiac arrhythmia syndromes. We established a denaturing high-performance liquid chromatography (DHPLC) protocol for rapid mutation screening of these genes, and reported mutations and variations identified by this method. We included 28 patients with Brugada syndrome, 4 with congenital long QT syndrome (LQTS), 11 with drug-induced LQTS, 4 with idiopathic ventricular fibrillation, and 50 normal volunteers. Polymerase chain reactions were performed to amplify the entire coding region of these genes. DHPLC was used to screen for heteroduplexes then DNA sequencing was performed. With this method, we identified the mutation(s) in all four patients with congenital LQTS (KCNQ1 A341V, KCNH2 N633D, KCNH2 2768Cdel and KCNE1 K70 N Y81C double mutations). We also identified the SCN5A A551T mutation in 1 of the 28 patients with Brugada syndrome. All the above-mentioned mutations were novel except KCNQ1 A341V. No mutations were identified in patients with drug-induced LQTS or idiopathic ventricular fibrillation. In total, 25 single nucleotide polymorphisms were identified, 10 of which were novel. In conclusion, DHPLC is a sensitive and rapid method for detection of cardiac sodium and potassium channel gene mutations.
authors.affiliation: Institute of Pharmacology, National Taiwan University Hospital, Taipei, Taiwan.
authors.forename: Ling-Ping
authors.initials: LP
authors.lastname: Lai
authors.affiliation: Department of Medical Genetics, National Taiwan University Hospital, Taipei, Taiwan.
authors.forename: Yi-Ning
authors.initials: YN
authors.lastname: Su
authors.affiliation: Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
authors.forename: Fu-Tien
authors.initials: FT
authors.lastname: Chiang
authors.affiliation: Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
authors.forename: Jyh-Ming
authors.initials: JM
authors.lastname: Juang
authors.affiliation: Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
authors.forename: Yen-Bin
authors.initials: YB
authors.lastname: Liu
authors.affiliation: Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
authors.forename: Yi-Lwun
authors.initials: YL
authors.lastname: Ho
authors.affiliation: Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
authors.forename: Wen-Jone
authors.initials: WJ
authors.lastname: Chen
authors.affiliation: Department of Medicine, Chang Gung Memorial Hospital, Taipei, Taiwan.
authors.forename: San-Jou
authors.initials: SJ
authors.lastname: Yeh
authors.affiliation: Department of Medicine, Chang Gung Memorial Hospital, Taipei, Taiwan.
authors.forename: Chun-Chieh
authors.initials: CC
authors.lastname: Wang
authors.affiliation: Department of Medicine, Chang Gung Memorial Hospital, Taipei, Taiwan.
authors.forename: Yu-Lin
authors.initials: YL
authors.lastname: Ko
authors.affiliation: Taichung Veterans General Hospital, School of Medicine, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Tsu-Juey
authors.initials: TJ
authors.lastname: Wu
authors.affiliation: Division of Cardiology and Cardiovascular Surgery, Institute of Medicine, Chung-Shan Medical University Hospital, Taichung, Taiwan.
authors.forename: Kwo-Chang
authors.initials: KC
authors.lastname: Ueng
authors.affiliation: Department of Internal Medicine, Poh-Ai Hospital, Lotung, Taiwan.
authors.forename: Meng-Huan
authors.initials: MH
authors.lastname: Lei
authors.affiliation: Department of Internal Medicine, Poh-Ai Hospital, Lotung, Taiwan.
authors.forename: Hsuan-Ming
authors.initials: HM
authors.lastname: Tsao
authors.affiliation: School of Medicine, Taipei Veterans General Hospital, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Shih-Ann
authors.initials: SA
authors.lastname: Chen
authors.affiliation: Cardiovascular Division, Department of Internal Medicine, Buddhist Dalin Tzu Chi General Hospital, Dalin, Taiwan.
authors.forename: Tin-Kwang
authors.initials: TK
authors.lastname: Lin
authors.affiliation: Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan.
authors.forename: Mei-Hwan
authors.initials: MH
authors.lastname: Wu
authors.affiliation: Department of Internal Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan.
authors.forename: Huey-Ming
authors.initials: HM
authors.lastname: Lo
authors.affiliation: Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
authors.forename: Shoei K Stephen
authors.initials: SKS
authors.lastname: Huang
authors.affiliation: Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan. jiunn@ha.mc.ntu.edu.tw.
authors.forename: Jiunn-Lee
authors.initials: JL
authors.lastname: Lin
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods; D017931:DNA Primers; D005820:Genetic Testing / Q000379:methods*; D006323:Heart Arrest / Q000235:genetics*; D006801:Humans; D008133:Long QT Syndrome / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D009206:Myocardium / Q000378:metabolism*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015221:Potassium Channels / Q000235:genetics*; D017422:Sequence Analysis, DNA; D015222:Sodium Channels / Q000235:genetics*; D013577:Syndrome; D013624:Taiwan
chemical_list: D017931:DNA Primers; D015221:Potassium Channels; D015222:Sodium Channels