MTHFR and MTRR gene polymorphism detection primer, probe, kit and application

Abstract

The invention provides an MTHFR and MTRR gene polymorphism detection primer and probe combination, which includes an MTHFR C677T, MTHFR A1298C and MTRR A66G gene polymorphism detection primer and probe combination. The invention further provides a detection kit for the MTHFR and MTRR gene polymorphism. The kit is prepared from the following components of a nucleic acid releasing agent, 2*PCR reaction mixture (including a thermally activated Taq DNA polymerase and a UDG enzyme), three sets of specific primers, three sets of specific probes and an internal standard system. The kit only needs tobriefly treat a 2 [mu]l blood sample at the room temperature to produce high-quality DNA for repeated detection, the detection specificity is good, the coincidence rate with a gold standard sequencingmethod is 100%, the sensitivity is high, accurate typing of 0.2ng genomic DNA can be achieved, the detection process is short in consumed time, the process from sample processing to detection resultscan be completed within 1 hour.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a primer, a probe, a kit for rapid detection of MTHFR and MTRR gene polymorphisms and applications thereof. Background technique [0002] Folic acid is a water-soluble B vitamin, an element necessary for the synthesis of nucleic acid, a substance necessary for cell growth and tissue repair, and an indispensable nutrient for embryonic development. Reduced activity of key enzymes in the folic acid metabolic pathway can lead to hypofolate and hyperhomocysteinemia, which is a major cause of birth defects in newborns, pregnancy-induced hypertension in pregnant women, spontaneous abortion, decreased sperm quality in men, H-type hypertension, One of the important risk factors for atherosclerosis and coronary heart disease. [0003] 5,10-methylenetetrahydrofolate reductase (MTHFR) and methionine synthase reductase (MTRR) are key enzymes that affect folic acid metabolism, and...

AI Technical Summary

Problems solved by technology

Among them, the DNA sequencing method is the gold standard for mutation detection, but this method has disadvantages such as complicated operation, long detection cycle, easy pollution, and high cost; the gene chip method is difficult to prepare chips, cumbersome to operate, and expensive to detect; the HRM method requires high equipment requirements , special analysis software needs to be configured, which limits the popularization of this method; the traditional fluorescent quantitative PCR method is widely used in clinical practice, but this method generally uses the Genotyping method for genotyping, which has certain requirements for the number of samples and the distribution of polymorphisms. When the sample size is small, the polymorphism of the sample cannot be accurately typed

In addition, the template DNA used in the above detection methods needs to go through cumbersome extraction and purification steps, which is not only time-consuming and costly, but also easily leads to nucleic acid loss and cross-contamination between samples

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