Primer in use for in vitro diagnosing GJB2 mutation of deaf gene of autosomal recessive inheritance in non-syndrome

Abstract

This invention provides primers for in vitro diagnosis of mutation of autosomal recessive nonsyndromic hearing loss gene GJB2. The primers can be used for completely amplifying the coding region and upstream or downstream important splicing sequences of GJB2 gene. This invention also provides a test kit for containing the primers and ApaI restriction endonuclease and their application for in vitro detection of 233-235delC mutation of the complete GJB2 coding region. The test kit can also be used for detecting the mutation sites of the GJB2 coding region through sequencing by the forward primer, and detecting deletion or insertion heterozygous mutation through sequencing by the reverse primer. The test kit is suitable for large-scale screening of autosomal recessive nonsyndromic hearing loss gene GJB2 mutation and genetic counseling.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a primer for in vitro diagnosis of mutations in the entire coding region of the non-syndromic autosomal recessive deafness gene GJB2 gene and 233-235delC hotspot mutations. The present invention also relates to the application of the primer in the preparation of a kit or similar product for the in vitro diagnosis of mutations in the entire coding region of the non-syndromic autosomal recessive deafness gene GJB2, particularly a kit or similar product containing the primer , which can be used for in vitro diagnosis of mutations in the entire coding region of the non-syndromic autosomal recessive deafness gene GJB2 gene and 233-235delC hotspot mutations. Background technique [0002] Currently, 1 / 1000-3 / 1000 children are born with profound deafness. More than half of childhood deafness is hereditary deafness, and most of them are autosomal recessive and non-syndromic d...

AI Technical Summary

Problems solved by technology

However, the above method is not suitable for screening and promotion due to its high cost, expensive equipment, complicated operation, high technical requirements, and time-consuming and labor-intensive use.

[0006] Since we found that the mutation hotspot of the Chinese GJB2 gene is at the 233-235 site, and deaf patients with this site mutation can confirm that their deafness is caused by the GJB2 gene mutation; after the deletion of base C in the 233-235 site, It leads to a change in the sequence of the 230-235 position, so compared with the wild-type sample, the sample with the delC mutation at the 230-235 position of the GJB2 gene cannot be digested by the endonuclease ApaI

Images