Fluorescent quantitative PCR detection kit for mitochondrion deafness A7445G mutation and application thereof

Abstract

The invention provides a fluorescent quantitative PCR detection kit for mitochondrion deafness A7445G mutation. The kit is prepared from quantitative PCR reaction liquid, a first positive control substance, a second positive control substance, a negative control substance, a specification and a box body, wherein the quantitative PCR reaction liquid is prepared from a PCR buffer solution, MgCl2, dNTPs, a heat-resistant DNA polymerase, an upstream amplification primer, a downstream amplification primer, a first fluorescent probe and a second fluorescent probe. According to the kit provided by the invention, two MGB probes are designed for the A being larger than G single-base mutation on the site 7445 of mtDNA; the nucleotide on the site 7445 of the deafness related mitochondrion is detected through fluorescent quantitative PCR to determine whether the A being larger than G mutation happens on the site; the kit is simple, quick, accurate and the like and can be applied to clinical diagnosis and control of mitochondrion A7445G mutation related deafness and also provides a basis for the prevention and treatment of mitochondrion deafness caused by A7445G 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 A7445G 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] Deafness-associated mtDNA tRNA gene mutations involve a total of 16 tRNAs, among which tRNA Ser(UCN) Gen...

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

Images