A Reverse Dot Blot Assay for the Screening of Twenty Mutations in Four Genes Associated with NSHL in a Chinese Population

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 May 16

PMID 28505178

Citations 6

Authors

Siping Li

Qi Peng

Shengyun Liao

Wenrui Li

Qiang Ma

Xiaomei Lu

Affiliations

Soon will be listed here.

Abstract

Background: Congenital deafness is one of the most distressing disorders affecting humanity and exhibits a high incidence worldwide. Most cases of congenital deafness in the Chinese population are caused by defects in a limited number of genes. A convenient and reliable method for detecting common deafness-related gene mutations in the Chinese population is required.

Methods: We developed a PCR-reverse dot blot (RDB) assay for screening 20 hotspot mutations of GJB2, GJB3, SLC26A4, and MT-RNR1, which are common non-syndromic hearing loss (NSHL)-associated genes in the Chinese population. The PCR-RDB assay consists of multiplex PCR amplifications of 10 fragments in the target sequence of the four above-mentioned genes in wild-type and mutant genomic DNA samples followed by hybridization to a test strip containing allele-specific oligonucleotide probes. We applied our method to a set of 225 neonates with deafness gene mutations and 30 normal neonates.

Results: The test was validated through direct sequencing in a blinded study with 100% concordance.

Conclusions: The results demonstrated that our reverse dot blot assay is a reliable and effective genetic screening method for identifying carriers and individuals with NSHL among the Chinese population.

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