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.
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.
Lai
Ling-Ping
LP
Institute of Pharmacology, National Taiwan University Hospital, Taipei, Taiwan.
Su
Yi-Ning
YN
Department of Medical Genetics, National Taiwan University Hospital, Taipei, Taiwan.
Chiang
Fu-Tien
FT
Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
Juang
Jyh-Ming
JM
Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
Liu
Yen-Bin
YB
Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
Ho
Yi-Lwun
YL
Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
Chen
Wen-Jone
WJ
Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
Yeh
San-Jou
SJ
Department of Medicine, Chang Gung Memorial Hospital, Taipei, Taiwan.
Wang
Chun-Chieh
CC
Department of Medicine, Chang Gung Memorial Hospital, Taipei, Taiwan.
Ko
Yu-Lin
YL
Department of Medicine, Chang Gung Memorial Hospital, Taipei, Taiwan.
Wu
Tsu-Juey
TJ
Taichung Veterans General Hospital, School of Medicine, National Yang-Ming University, Taipei, Taiwan.
Ueng
Kwo-Chang
KC
Division of Cardiology and Cardiovascular Surgery, Institute of Medicine, Chung-Shan Medical University Hospital, Taichung, Taiwan.
Lei
Meng-Huan
MH
Department of Internal Medicine, Poh-Ai Hospital, Lotung, Taiwan.
Tsao
Hsuan-Ming
HM
Department of Internal Medicine, Poh-Ai Hospital, Lotung, Taiwan.
Chen
Shih-Ann
SA
School of Medicine, Taipei Veterans General Hospital, National Yang-Ming University, Taipei, Taiwan.
Lin
Tin-Kwang
TK
Cardiovascular Division, Department of Internal Medicine, Buddhist Dalin Tzu Chi General Hospital, Dalin, Taiwan.
Wu
Mei-Hwan
MH
Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan.
Lo
Huey-Ming
HM
Department of Internal Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan.
Huang
Shoei K Stephen
SKS
Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan.
Lin
Jiunn-Lee
JL
Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S Road, Taipei, Taiwan. jiunn@ha.mc.ntu.edu.tw.
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
D017931:DNA Primers; D015221:Potassium Channels; D015222:Sodium Channels