Kit and detection method for polymorphism detection of methylenetetrahydrofolate reductase gene

Abstract

The invention relates to a kit and a detection method for polymorphism detection of a methylenetetrahydrofolate reductase gene. The kit comprises a nucleic acid extraction reagent and a nucleic acid detection reagent, wherein the nucleic acid detection reagent comprises fluorescent probe-containing PCR reaction liquid, a negative control and a positive control; the nucleic acid extraction reagent is a DNA extract; the PCR reaction liquid comprises DNA polymerase, UNG enzyme, a primer, a probe and Mg<2+>; the probe is a molecular beacon probe; and the gene polymorphic site of MTHFR is a C677T polymorphic site. The detection method using the kit is easy and fast to operate, the use of consumables is reduced so as to reduce the detection cost, the detection result is accurate, and the occurrence of false positive result is reduced; by adding the negative control and positive control in the amplification step, the judgment of false positive and false negative can be effectively improved, and the detection precision is higher; since the hairpin structure of the molecular beacon probe is opened with certain force, the specificity to the mutant C677T polymorphic site at the measuring point is higher.

Description

technical field [0001] The invention relates to the field of medical reagents, in particular to a gene diagnosis kit, in particular to a kit and a detection method for detecting the polymorphism of the methylenetetrahydrofolate reductase gene. Background technique [0002] Methylenetetrahydrofolate reductase gene (MTHFR) is a one-carbon unit metabolizing enzyme, present in animal liver. It is a key enzyme in the folic acid methylation process. Its main function is to reduce 5,10-tetrahydromethylfolate to 5-methyltetrahydrofolate, provide methyl for the conversion of homocysteine ​​to methionine, and It is DNA, participates in the synthesis of purine and pyrimidine in the form of one-carbon units, and provides methyl for DNA, RNA and protein methylation, affecting DNA metabolism. [0003] Methyl donor for a variety of biological substrates including proteins and neurotransmitter factors. Methionine is an essential amino acid in the human body, and the synthesis of methionin...

AI Technical Summary

Problems solved by technology

[0023] Due to the lack of folic acid and the impact of homocysteine ​​on many related diseases, the detection of MTHFR gene polymorphism is particularly important. However, the commonly used clinical detection methods have some defects and lack of predictability. In order to predict the homocysteine ​​earlier To prevent and intervene the occurrence and progression of diseases associated with homocysteine, it is necessary to predict the level of homocysteine ​​as early as possible

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