Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis

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 ana...

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Autores principales: Wang, Xudong, Hong, Yongjun, Cai, Peihong, Tang, Ning, Chen, Ying, Yan, Tizhen, Liu, Yinghua, Huang, Qiuying, Li, Qingge
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5320477/
https://www.ncbi.nlm.nih.gov/pubmed/28225033
http://dx.doi.org/10.1038/srep42894
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author Wang, Xudong
Hong, Yongjun
Cai, Peihong
Tang, Ning
Chen, Ying
Yan, Tizhen
Liu, Yinghua
Huang, Qiuying
Li, Qingge
author_facet Wang, Xudong
Hong, Yongjun
Cai, Peihong
Tang, Ning
Chen, Ying
Yan, Tizhen
Liu, Yinghua
Huang, Qiuying
Li, Qingge
author_sort Wang, Xudong
collection PubMed
description 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.
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spelling pubmed-53204772017-02-24 Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis Wang, Xudong Hong, Yongjun Cai, Peihong Tang, Ning Chen, Ying Yan, Tizhen Liu, Yinghua Huang, Qiuying Li, Qingge Sci Rep Article 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. Nature Publishing Group 2017-02-22 /pmc/articles/PMC5320477/ /pubmed/28225033 http://dx.doi.org/10.1038/srep42894 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Wang, Xudong
Hong, Yongjun
Cai, Peihong
Tang, Ning
Chen, Ying
Yan, Tizhen
Liu, Yinghua
Huang, Qiuying
Li, Qingge
Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis
title Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis
title_full Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis
title_fullStr Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis
title_full_unstemmed Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis
title_short Rapid and Reliable Detection of Nonsyndromic Hearing Loss Mutations by Multicolor Melting Curve Analysis
title_sort rapid and reliable detection of nonsyndromic hearing loss mutations by multicolor melting curve analysis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5320477/
https://www.ncbi.nlm.nih.gov/pubmed/28225033
http://dx.doi.org/10.1038/srep42894
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