Fluorescent quantitative PCR (Polymerase Chain Reaction) detection kit for mutation A1555G 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 A1555G 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 A>G mono-base mutation of a 1555th locus of mtDNA, and whether the locus is subjected to A>G mutation or not is determined through detecting 1555th 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 A1555G and can also be used for providing a foundation for preventing and treating the mitochondrial deafness induced by the mutation A1555G.

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 A1555G mutation and its application in the detection of mitochondrial deafness A1555G mutation. 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] The A1555G mutation located on mtDNA 12S rRNA ...

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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