title: Rapid identification of aminoglycoside-induced deafness gene mutations using multiplex real-time polymerase chain reaction.
abstract: BACKGROUND
  Exposure to aminoglycoside antibiotics can induce ototoxicity in genetically susceptible individuals carrying certain mitochondrial DNA (mtDNA) mutations (C1494T and A1555G), resulting in hearing loss. So, a rapid diagnostic approach is needed to accurately identify subjects carrying such gene mutations.
  
  
  METHODS
  In the present study, we describe a rapid and reliable four-color, real-time quantitative polymerase chain reaction (qPCR) assay for simultaneously detecting two mtDNA 12S rRNA gene variants, A1555G and C1494T, which are prevalent in the Han Chinese population. This multiplex assay incorporates three allele-specific TaqMan probes labeled with different fluorophores in a single reaction, providing high genotyping accuracy for clinical blood samples.
  
  
  RESULTS
  Tests with C1494T, A1555G and wild-type DNA exhibited high sensitivity, specificity, reproducibility and accuracy of discriminating mutations from wild-type.
  
  
  CONCLUSIONS
  This study shows that this simple and inexpensive method can be used for routine molecular diagnostics and potentially for large-scale genetic screening.
authors.affiliation: Department of Otolaryngology, PLA General Hospital, Do. 28 Fuxing Rode, Beijing 100853, People's Republic of China.
authors.forename: Shasha
authors.initials: S
authors.lastname: Huang
authors.affiliation: Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, P. R. China; National Engineering Research Center for Beijing Biochip Technology, Beijing 102206, P. R. China; CapitaBio Corporation, Beijing 102206, P. R. China.
authors.forename: Guangxin
authors.initials: G
authors.lastname: Xiang
authors.affiliation: Department of Otolaryngology, PLA General Hospital, Do. 28 Fuxing Rode, Beijing 100853, People's Republic of China.
authors.forename: Dongyang
authors.initials: D
authors.lastname: Kang
authors.affiliation: Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, P. R. China; National Engineering Research Center for Beijing Biochip Technology, Beijing 102206, P. R. China.
authors.forename: Chen
authors.initials: C
authors.lastname: Wang
authors.affiliation: Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, P. R. China; National Engineering Research Center for Beijing Biochip Technology, Beijing 102206, P. R. China.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Kong
authors.affiliation: Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, P. R. China; National Engineering Research Center for Beijing Biochip Technology, Beijing 102206, P. R. China.
authors.forename: Xun
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, P. R. China; National Engineering Research Center for Beijing Biochip Technology, Beijing 102206, P. R. China.
authors.forename: Shujian
authors.initials: S
authors.lastname: Liang
authors.affiliation: Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, P. R. China; National Engineering Research Center for Beijing Biochip Technology, Beijing 102206, P. R. China; Medical Systems Biology Research Center, Department of Biomedical Engineering, Tsinghua University School of Medicine, Beijing 100084, P. R. China.
authors.forename: Keith
authors.initials: K
authors.lastname: Mitchelson
authors.affiliation: Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, P. R. China; National Engineering Research Center for Beijing Biochip Technology, Beijing 102206, P. R. China; CapitaBio Corporation, Beijing 102206, P. R. China; Medical Systems Biology Research Center, Department of Biomedical Engineering, Tsinghua University School of Medicine, Beijing 100084, P. R. China. Electronic address: wlxing@tsinghua.edu.cn.
authors.forename: Wanli
authors.initials: W
authors.lastname: Xing
authors.affiliation: Department of Otolaryngology, PLA General Hospital, Do. 28 Fuxing Rode, Beijing 100853, People's Republic of China. Electronic address: daipu301@vip.sina.com.
authors.forename: Pu
authors.initials: P
authors.lastname: Dai
mesh_terms: D004272:DNA, Mitochondrial / Q000235:genetics*; D005819:Genetic Markers; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods*; D006319:Hearing Loss, Sensorineural / Q000139:chemically induced* / Q000235:genetics; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D009154:Mutation; D012335:RNA, Ribosomal / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004272:DNA, Mitochondrial; D005819:Genetic Markers; D012335:RNA, Ribosomal; C075526:RNA, ribosomal, 12S
keywords: Hearing loss; Maternal inheritance; Mitochondrial DNA; Real-time quantitative polymerase chain reaction; Sensorineural