A reverse dot blot assay for the screening of twenty mutations in four genes associated with NSHL in a Chinese population

PloS One
Siping LiXiaomei Lu

Abstract

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

References

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Oct 18, 2016·Genetic Testing and Molecular Biomarkers·Qi PengXiaomei Lu

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Citations

Jun 7, 2019·Genetics in Medicine : Official Journal of the American College of Medical Genetics·A Eliot ShearerUNKNOWN Newborn Hearing Screening Working Group of the National Coordinating Center for the Regional Genetics Networks
Mar 8, 2019·Scientific Reports·Mavys Tabraue-ChávezJuan José Díaz-Mochón

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Methods Mentioned

BETA
PCR
electrophoresis
dot blot
dot
genotyping

Software Mentioned

MassARRAY

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