Hereditary deafness gene mutation detection kit

Abstract

The invention relates to a hereditary deafness gene mutation detection kit. The hereditary deafness gene mutation detection kit comprises sequencing primers, wherein the sequencing primers comprise sequences as follows: SEQ ID NO:1-36. The hereditary deafness gene mutation detection kit has the advantages of multiple detection site, wide coverage range, high positive rate, high clinical usage value, high sensitivity, high accuracy, high detection specificity, high repeatability and good detection stability and can be applied to premarital screening, prenatal diagnosis and hereditary deafness gene site screening and detection for fetuses during pregnancy and deaf people, so that the birth of deaf children and occurrence of tardive deafness can be greatly avoided, economic burdens on the society and families can be reduced, and the good birth and the good care can be realized.

Description

technical field [0001] The invention relates to gene sequencing technology, in particular to a genetic mutation detection kit for deafness. Background technique [0002] Deafness is one of the most common sensory impairment diseases worldwide. Hereditary deafness can be divided into non-syndromic deafness and syndromic deafness. According to the second national sample survey of disabled people, there are about 20.04 million people with hearing disabilities in my country, accounting for 24.16% of the total number of disabled people; about 1.27 million people with speech disabilities caused by deafness, accounting for 1.53% of the total number of disabled people. The incidence of congenital deafness in newborns is about 1 / 1000, and more than half of the congenital deafness is related to genetics. [0003] In non-syndromic deafness, GJB2, SLC26A4 and mitochondrial DNA are the most common causative genes in my country. The GJB2 gene encodes connexin 26, which is involved in in...

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Problems solved by technology

[0005] Microarray chip detection requires whole blood DNA extraction (30min), PCR amplification (over 2h), product hybridization analysis (over 3h), etc. The detection time is long (over 6h), and the operation is cumbersome, which cannot meet the clinical requirements for deafness. The need for a large number of genetic screening; due to too many operating procedures, it is easy to cause sample cross-contamination; secondly, the recognition rate of a single base is relatively low, and false negatives and false positives are prone to occur, which affects the accuracy of the results; in addition, micro The cost of array chips is too high, requiring special chip scanning instruments, and the detection cost is high; the amplification efficiency of ordinary ARMS-PCR products is only 1-10% of normal amplification, so the amplification system lacks stability; four-primer ARMS-PCR One reaction can only detect one mutation site

The common disadvantages of common PCR products are: the genotype needs to be judged by the size of the band when interpreting the results, which lacks intuition

Gel electrophoresis detection results are prone to PCR cross-contamination, low detection sensitivity and time-consuming; using Taqman probes for SNP mutation detection requires 2 probes for each site detected, and has high requirements for probe specificity

The cost of probe synthesis is high, and there are requirements for the number of channels of the fluorescent PCR instrument, which is not conducive to popularization; the detection of one tube by the fluorescent melting curve method has few detection sites, and is limited by the channels of fluorescent dyes, which cannot meet the detection of multiple genes and multiple sites of deafness; PCR-RFLP detection technology is cumbersome to operate, requires enzyme digestion reaction, takes a long time to react, and requires gel electrophoresis detection, which is prone to cross-contamination of samples

Restriction sites are easily affected by gene variation; the multiplex PCR primer system is complex and difficult to design, and multiple pairs of primers will interfere with each other and competitively inhibit

Therefore, the more detection sites, the more primers required, the more difficult it is to optimize the PCR system. At the same time, due to the limitation of the fluorescence channel of the capillary electrophoresis instrument, it is difficult to promote the product. Generally, the maximum number of detection sites does not exceed 20;

[0006] The common disadvantage of existing products is that there are few detection sites. They can only detect hotspot mutations in hereditary deafness genes, but cannot detect many rare sites. However, there are many deafness-causing genes and mutation sites. , simple, wide detection range requirements

There is a possibility of missed detection in clinical application, and it is impossible to further explain the cause of the patient, which reduces the value of clinical use

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