Primer pair, probe set and kit for detecting mitochondrial obesity gene mutation

Abstract

The invention belongs to the technical field of biological detection, and discloses a primer pair, a probe set and a kit for detecting mitochondrial obesity gene mutation. The kit takes human DNA as a formwork, fluorescence quantitative PCR amplification is carried out to obtain an amplification curve, and a mitochondrial tRNA Leu (UUR) 3243A more than G mutation site can be quickly, accurately and sensitively detected. The kit is further provided with a wild type site probe, and the relative ratio of a mutant type to a wild type is obtained through Ct values of the mutant type and the wild type probe. The blank of clinically detecting the tRNA Leu (UUR) 3243A more than G mutation site is filled, and timely auxiliary data is provided for subsequent treatment of related diseases. The kit is good in repeatability and high in precision. The kit is short in detection period, detection can be completed within 45 minutes at the soonest, the detection time is saved, the efficiency is improved, and the kit is an efficient auxiliary detection means. In the application of non-diagnosis purposes, detection can be completed only by mixing the kit and a collected sample through a fluorescence quantitative PCR instrument, the operation requirement is simple, and the clinical popularization performance is high.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a primer pair, a probe set and a kit for detecting mitochondrial obesity gene mutations. Background technique [0002] Mitochondrial disease refers to the abnormality of mitochondrial oxidative respiratory chain-related proteins due to mutations or deletions of mitochondrial DNA and nuclear DNA, which in turn leads to multiple organ dysfunction syndrome that hinders ATP synthesis. The types of mitochondrial gene mutations include base mutations, insertion and deletion mutations, and mtDNA copy number mutations. tRNA Leu(UUR)3243A>G is the most common mitochondrial gene mutation, which is usually maternally inherited, but sporadic mutations can occasionally lead to A series of clinical symptoms ranging from mild to fatal, one of which is diabetes. [0003] The incidence of tRNA Leu(UUR)3243A>G mutation in diabetic patients can reach more than 50%, a...

AI Technical Summary

Problems solved by technology

At present, relevant research at home and abroad is based on the distribution of clinical phenotypes of their respective cohorts of patients, limited scope, racial differences, and the coverage of each study is not completely the same

The detection method of tRNA Leu(UUR)3243A>G mutation has the traditional PCR digestion method, which is complicated to operate and has cumbersome steps

The method of high-throughput sequencing can also be used, but it is expensive; while the method of fluorescent quantitative PCR has the characteristics of simple operation, convenience and quickness

[0005] Related technologies disclose the use of fluorescent quantitative PCR method to quantitatively detect the mutation site of tRNA Leu(UUR)3243A>G, but this method uses a detection system with a long detection cycle, which is not conducive to providing clinical detection results in time; on the other hand, this method Although the invention can carry out relative quantification of the mutation site, it does not carry out relative quantification of the wild type and the mutant type. Only by clarifying the relative ratio of the wild type and the mutant type can it provide better guidance for the formulation of subsequent intervention plans

[0006] At present, there is no CFDA-approved kit for clinical detection of tRNA Leu(UUR)3243A>G mutation in China

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