Document Detail

Mutation analysis of SLC26A4 for Pendred syndrome and nonsyndromic hearing loss by high-resolution melting.
MedLine Citation:
PMID:  21704276     Owner:  NLM     Status:  MEDLINE    
Pendred syndrome and DFNB4 (autosomal recessive nonsyndromic congenital deafness, locus 4) are associated with autosomal recessive congenital sensorineural hearing loss and mutations in the SLC26A4 gene. Extensive allelic heterogeneity, however, necessitates analysis of all exons and splice sites to identify mutations for individual patients. Although Sanger sequencing is the gold standard for mutation detection, screening methods supplemented with targeted sequencing can provide a cost-effective alternative. One such method, denaturing high-performance liquid chromatography, was developed for clinical mutation detection in SLC26A4. However, this method inherently cannot distinguish homozygous changes from wild-type sequences. High-resolution melting (HRM), on the other hand, can detect heterozygous and homozygous changes cost-effectively, without any post-PCR modifications. We developed a closed-tube HRM mutation detection method specific for SLC26A4 that can be used in the clinical diagnostic setting. Twenty-eight primer pairs were designed to cover all 21 SLC26A4 exons and splice junction sequences. Using the resulting amplicons, initial HRM analysis detected all 45 variants previously identified by sequencing. Subsequently, a 384-well plate format was designed for up to three patient samples per run. Blinded HRM testing on these plates of patient samples collected over 1 year in a clinical diagnostic laboratory accurately detected all variants identified by sequencing. In conclusion, HRM with targeted sequencing is a reliable, simple, and cost-effective method for SLC26A4 mutation screening and detection.
Neng Chen; Lisbeth Tranebjærg; Nanna Dahl Rendtorff; Iris Schrijver
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-04-29
Journal Detail:
Title:  The Journal of molecular diagnostics : JMD     Volume:  13     ISSN:  1943-7811     ISO Abbreviation:  J Mol Diagn     Publication Date:  2011 Jul 
Date Detail:
Created Date:  2011-06-27     Completed Date:  2011-10-17     Revised Date:  2013-06-28    
Medline Journal Info:
Nlm Unique ID:  100893612     Medline TA:  J Mol Diagn     Country:  United States    
Other Details:
Languages:  eng     Pagination:  416-26     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
Department of Pathology, Stanford University School of Medicine, Stanford, California, USA.
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MeSH Terms
DNA Mutational Analysis
Goiter, Nodular / diagnosis,  genetics*
Hearing Loss, Sensorineural / diagnosis,  genetics*
Membrane Transport Proteins / genetics*
Molecular Diagnostic Techniques*
Reproducibility of Results
Transition Temperature
Reg. No./Substance:
0/Membrane Transport Proteins; 0/SLC26A4 protein, human

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