Standard substance and kit for detecting mitochondrial A3243G heterozygous mutation rate and detection method

Abstract

The invention discloses a standard substance and a kit for detecting mitochondrial A3243G heterozygous mutation rate and a detection method. The standard substance for detecting mitochondrial A3243G heterozygous mutation rate provided by the invention comprises: (1) a vector containing a mitochondrial DNA fragment with 3243 locus being a wild type A; and (2) a vector containing a mitochondrial DNA fragment with 3243 locus being a mutant G. The plasmid molecule standard substance, the detection method and a regression equation used for detecting data correction provided by the invention can conveniently and accurately detect A3243G heterozygous mutation rate in the mitochondrial DNA, and have advantages of high efficiency and sensitivity. In addition, the plasmid molecule standard substance, the detection method and the regression equation can accurately and quantificationally detect heterozygous mutation with mutation rate lower than 5%, which is a unique advantage unachievable by other heterozygous mutation detection means. The invention also has good reference meaning for quantitative determination of other heterozygous mutation in scientific research and production practice.

Description

technical field

[0001] The invention relates to the field of biomedical engineering, in particular to a standard product, a kit and a detection method for detecting the mitochondrial A3243G heterozygous mutation rate. Background technique

[0002] The A3243G mutation is the most common pathogenic mutation on mitochondrial genomic DNA (mito-chondrial DNA, mtDNA), which often exists in a heterozygous form, that is, there are both wild-type mtDNA and mutant mtDNA in a cell. A3243G mutation is one of the pathogenic factors of premature diabetes and neurological deafness, and the A3243G mutation rate is related to the severity of clinical symptoms. Therefore, the accurate quantitative detection of the A3243G mutation is of great significance. The mtDNA3243 site is located between 16S rRNA and tRNA Leu At the junction of the (UUR) gene, the study of the A / G mutation at the mtDNA3243 site found that the A base is located on the dihydrourinophilic loop of the secondary structure o...

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