Fluorescent quantitative PCR (Polymerase Chain Reaction) detection kit for mutation T12201C of mitochondrial deafness and application of fluorescent quantitative PCR detection kit

Abstract

The invention provides a fluorescent quantitative PCR (Polymerase Chain Reaction) detection kit for common mutation T12201C of mitochondrial deafness. The kit comprises quantitative PCR reaction solutions, a first positive control, a second positive control, a negative control, a description and a box body, wherein the quantitative PCR reaction solutions contain a PCR buffer solution, MgCl2, dNTPs, 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 T>C mono-base mutation of a 12201th locus of mtDNA, and whether the locus is subjected to T>C mutation or not is determined through detecting 12201th nucleotide of a mitochondrion correlated to deafness by fluorescent quantitative PCR, so that the kit has the advantages of simplicity, convenience, rapidness, accuracy and the like, can be applied to the clinical diagnosis and control of deafness related to mitochondrial mutation T12201C and can also be used for providing a foundation for preventing and treating the mitochondrial deafness induced by the mutation T12201C.

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 T12201C 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-related mtDNA tRNA gene mutations involve 16 tRNAs, and the T12201C mutation is located in the ...

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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