Specific primer and test kit for detecting pathogenic variation of deafness gene based on multiplex PCR and high-throughput sequencing technology and application

Abstract

The invention relates to a specific primer and a test kit for detecting pathogenic variation of a deafness gene based on multiplex PCR and a high-throughput sequencing technology and application. Mosthereditary deafness can be subjected to high-efficiency, low-cost and comprehensive diagnosis; the specific primer and the test kit can also be used for newborn screening and early detection early treatment and can reduce matte caused by deafness, and the technical scheme for solving the problem is as follows: the multiplex PCR specific primer comprises 34 pairs of specific primers which are divided into two Pools, and Pool-1 comprises 18 pairs of primers, wherein the nucleic acid sequences are shown as SEQ ID NO: 1 to SEQ ID NO: 36; pool-2 comprises 16 pairs of primers, and the nucleotide sequence is shown as SEQ ID NO.37-68. According to the specific primer, the detection efficiency and accuracy can be effectively improved, the cost is reduced, the operation steps are simplified, and the specific primer is an innovation of specific primers for detecting the pathogenic variation of the deafness gene.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a specific primer, kit and application for detecting pathogenic variants of deafness genes based on multiplex PCR and high-throughput sequencing technology. Background technique [0002] Deafness is one of the most common disabilities, which can seriously affect human cognition and communication. There are as many as 360 million hearing-impaired people worldwide (WHO, 2017). About 1 in 500 newborns is affected by hearing loss. In China, there are about 800,000 hearing-impaired children under the age of 7, and this number is increasing at a rate of 30,000 cases every year. Non-syndromic or syndromic deafness associated with genetic factors accounts for about 60%. Non-synthetic deafness usually refers to deafness without other clinical symptoms, accounting for 70% of hereditary deafness. About 80% of sensorineural deafness is autosomal recessive (AR), besides, close to 20% of deafn...

AI Technical Summary

Problems solved by technology

[0009] One-generation sequencing is the traditional method for genetic testing of hereditary deafness, and is the gold standard for genetic testing; however, this technology is time-consuming, laborious, low-throughput and high-priced, which limits its clinical application

The microarray chip diagnostic technology combined with deafness genes can quickly obtain results, but the method contains a limited number of genes and loci, and cannot detect rare variants carried by patients, resulting in many patients being undiagnosed

Denaturing high performance liquid chromatography has the characteristics of high automation and low detection cost, and can detect multiple mutation sites at the same time, but its technical requirements are high and special instruments are required, which limits its clinical application, and is now mostly used in laboratory detection

Time-of-flight mass spectrometry has a high throughput. Each instrument can detect 3000 samples per day. It can detect many sites and is efficient and accurate. However, it requires high personnel skills and the purity of the test samples, and the instrument is expensive, so it is not suitable for Generally carried out by medical institutions, which limits its wide application

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