Kit for screening deaf gene of Chinese populations, and use method thereof

Abstract

The invention discloses a kit for one-time qualitative screening of common seven mutational sites of deaf gene of Chinese populations, and a use method thereof. Seven mutational hotspots which comprise delC of a 235th site of GJB2 gene, delAT of 299-300th sites of the GJB2 gene, A-G mutation of a 2168th site of SLC26A4 gene, A-G mutation of an IVS7-2th site of the SLC26A4 gene, A-G mutation of a 1555th site of mitochondrial DNA gene, A-G mutation of a 3243th site of the mitochondrial DNA gene, and A-G mutation of a 7445th site of the mitochondrial DNA gene are treated as detection objects, application primers and extension primers are respectively designed for the mutation of each site, multiple PCR amplification and markup extension are simultaneously carried out on each target site, and genotypes of above seven sites can be obtained at once through capillary electrophoretic analysis. The screening method and the kit of the invention have advantages of convenient use, simple operation, low cost, high flux, and direct and reliable detection result, and are suitable for large scale screening of the deaf gene mutation of the Chinese populations.

Description

technical field [0001] The invention relates to the field of gene mutation detection in the technical field of genetic engineering, in particular to a kit for screening deafness genes in Chinese population and a method for using the same. Background technique [0002] Deafness is a common condition that seriously affects quality of life and communication. There are more than 700 million people in the world suffering from moderate or above hearing loss. Among the 61 million people with disabilities in my country, 27 million are hearing and language disabled, and the number of newborn deaf children is increasing at a rate of 30,000 per year. Among them, hereditary NSHL ( nonsyndromic hearing loss, non-syndromic hearing loss) accounts for about 40%. So far, 28 autosomal recessive genes, 22 autosomal dominant genes, and 1 X-linked gene related to non-syndromic deafness have been cloned, and there are a large number of deafness mutation sites in each gene, Among them, the mutati...

AI Technical Summary

Problems solved by technology

[0008] At present, traditional gene mutation detection methods include single strand conformation polymorphism (Single Strand Conformation Polymorphism, SSCP), PCR-restriction fragment length polymorphism (Restriction Fragment Length Polymorphism, RFLP), denaturing high performance liquid chromatography (Denaturing High Performance Liquid Chromatography, DHPLC), direct sequencing, gene chips, etc. These methods either have limited detection capabilities, or low throughput, or are time-consuming and labor-intensive, and require expensive equipment and consumables

More importantly, it is difficult to perform high-throughput detection of multiple mutation sites of different genes at the same time in these methods except for gene chips, and the gene chip technology platform is expensive, requires high quality equipment and personnel, and is difficult to be used in the majority of medical institutions in my country. Therefore, it is necessary to create a high-throughput, high-efficiency, low-cost deafness gene mutation screening method to achieve rapid clinical detection or large-scale population screening

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