Hot Mutations Detection Of The Deafness By Using Multiple Single Nucleotide Primer Extension

Background:Lesions in external ear, middle ear, inner ear structures and all levels of the central nervous system in the auditory system, leading to listen dysfunction, thereby showing various degree of hearing loss, referred to as deafness. China has a large number of hearing disabilities. About30,000deaf children are born each year. The prevalence is1per500-1000. Approximately60%of deafness is genetic. More than70%of hereditary hearing loss is nonsyndromic while syndromic hearing impairment may account for up to30%. To date, about136nonsyndromic hearing impairment locus have been targeted, and88genes have been cloned. Despite the fact that deafness is a highly heterogeneous disorder, most nonsyndromic hearing impairment is caused by a few hot mutations in several specific genes, making genetic deafness screening become possible. The traditional diagnostic methods include restriction fragment length polymorphism, restriction endonuclease fingerprinting-single strand conformation polymorphism, denaturing performance liquid chromatography, Taqman MGB probe real time fluorescent quantitative, multiple polymerase chain reaction followed by Sanger sequencing, high resolution melting. However, these methods are difficult to simultaneously detect a plurality of different gene mutations. Microarray technology is expensive, though it can simultaneously detecte a plenty of genes’mutations. Therefore, it’s necessary to establish a reliable, economic and rapid method which could detect multiple mutations simultaneously.Objective:This study is aimed to simultaneously detect12hot mutations of the nonsyndromic hearing impairment probands in GJB2, GJB3, SLC26A4and12SrRNA by using multiple single nucleotide primer extension, and expect to establish a reliable, economic and rapid molecular genetic method for detecting the nonsyndromic hearing impairment.Methods:Multiple single nucleotide primer extension was used to simultaneously detect12hot mutations in GJB2, GJB3, SLC26A4and12SrRNA for212individuals, including96nonsyndromic hearing impairment probands,66family members, and50normal controls. Sanger sequencing was taken for confirmation.Results:1. The results made by the assay of multiple single nucleotide primer extension in the212individuals were consistent with Sanger sequencing, the accuracy of this assay was100%.2.41.67%of the96nonsyndromic hearing impairment probands was found to have pathogenic mutation by using multiple single nucleotide primer extension, while only31.25%was found by taking single technology.3. In this study, the carrier rate of nonsyndromic hearing impairment in normal persons was2%, and was consistent with previous studies.Conclusion:A reliable, economic and rapid method which could simultaneously detect12hot mutations in GJB2, GJB3, SLC26A4and12SrRNA was established.