Fluorescent quantitative PCR detection kit for mitochondrial deafness C1494T mutation and application thereof

Abstract

The invention provides a fluorescent quantitative PCR detection kit for mitochondrial deafness C1494T common mutation C1494T. The fluorescent quantitative PCR detection kit comprises a quantitative PCR reaction liquid, a first positive reference substance, a second positive reference substance, a negative reference substance, a direction and a case body, wherein the quantitative PCR reaction liquid contains a PCR buffer solution, MgCl2, dNTPs, heat-proof DNA polymerase, an upstream amplimer, a downstream amplimer, a first fluorescent probe and a second fluorescent probe. Two MGB probes are designed with respective to C>T single base mutation of the 1494 site of mtDNA, the deafness-related mitochondrial 1555 site nucleotide is detected through fluorescent quantitation PCR, and whether the site is subjected to C>T mutation or not is determined. The fluorescent quantitative PCR detection kit has the advantages of being simple, convenient, quick and accurate, can be applied to clinical diagnosis and treatment of mitochondrial C1494T mutation-related deafness and provides references for prevention and treatment of mitochondrial deafness caused by C1494T mutation.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic engineering, and specifically relates to a fluorescent quantitative PCR detection kit for mitochondrial deafness C1494T mutation and its application. Background technique [0002] Deafness is a public health problem of global concern, which is caused by genetic and environmental factors, and more than 50% of deafness patients are caused by genetic factors. In hereditary deafness, 30% are syndromic deafness and 70% are non-syndromic deafness. The inheritance mode of deafness can show autosomal dominant inheritance, autosomal recessive inheritance, X-linked inheritance and maternal inheritance. Mitochondrial DNA (mtDNA) mutation is one of the important causes of maternal hereditary deafness, and the incidence of hereditary deafness is The rate is about 1% to 2%. [0003] Clear literature has shown that the C1494T mutation located in mtDNA 12S rRNA is closely related to amino...

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problem of incomplete fluorescence quenching of traditional Taqman probes for fluorescent quantitative PCR, ABI Company of the United States introduced a new Taqman probe——MGB probe in 2000. Its 3' end uses a non-fluorescent quenching group. It does not emit light after absorbing the energy of the reporter group, which greatly reduces the interference of the background signal

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