title: Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis.
abstract: Hearing loss is a common birth defect worldwide. The GJB2, SLC26A4, MT-RNR1 and MT-TS1 genes have been reported as major pathogenic genes in nonsyndromic hearing loss. Early genetic screening is recommended to minimize the incidence of hearing loss. We hereby described a multicolor melting curve analysis (MMCA)-based assay for simultaneous detection of 12 prevalent nonsyndromic hearing loss-related mutations. The three-reaction assay could process 30 samples within 2.5 h in a single run on a 96-well thermocycler. Allelic types of each mutation could be reproducibly obtained from 10 pg ~100 ng genomic DNA per reaction. For the mitochondrial mutations, 10% ~ 20% heteroplasmic mutations could be detected. A comparison study using 501 clinical samples showed that the MMCA assay had 100% concordance with both SNaPshot minisequencing and Sanger sequencing. We concluded that the MMCA assay is a rapid, convenient and cost-effective method for detecting the common mutations, and can be expectedly a reliable tool in preliminary screening of nonsyndromic hearing loss in the Chinese Han population.
authors.affiliation: State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Engineering Research Centre of Molecular Diagnostics, Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China.
authors.forename: Xudong
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Otorhinolaryngology, Zhongshan Hospital of Xiamen, Xiamen University, Xiamen, Fujian, 361004, China.
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Hong
authors.affiliation: State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Engineering Research Centre of Molecular Diagnostics, Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China.
authors.forename: Peihong
authors.initials: P
authors.lastname: Cai
authors.affiliation: Department of Medical Genetics, Liuzhou Key Laboratory of Birth Defects Prevention and Control, Liuzhou Maternal and Child Health Hospital, Liuzhou, Guangxi 545001, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Tang
authors.affiliation: Nanjing Medical University Affiliated Wuxi Maternity and Child Health Care Hospital, Jiangsu 215002, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Medical Genetics, Liuzhou Key Laboratory of Birth Defects Prevention and Control, Liuzhou Maternal and Child Health Hospital, Liuzhou, Guangxi 545001, China.
authors.forename: Tizhen
authors.initials: T
authors.lastname: Yan
authors.affiliation: Department of Neonatology, Central Lab Suzhou Hospital Affiliated to Nanjing Medical University, Suzhou, Jiangsu 215002, China.
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Engineering Research Centre of Molecular Diagnostics, Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China.
authors.forename: Qiuying
authors.initials: Q
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Engineering Research Centre of Molecular Diagnostics, Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China.
authors.forename: Qingge
authors.initials: Q
authors.lastname: Li
mesh_terms: D003638:Deafness / Q000235:genetics*; D042241:Early Diagnosis; D005260:Female; D020022:Genetic Predisposition to Disease; D006801:Humans; D008297:Male; D008928:Mitochondria / Q000235:genetics; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D044366:Transition Temperature