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SNP detecting kit for methionine-folic acid metabolism key enzymes MTHFR and use method thereof

A detection kit and folic acid metabolism technology, applied in the field of medical detection, can solve the problems of unsuitability for rapid clinical detection, difficult clinical promotion, and long operation time, and achieve the effects of shortening detection time, easy judgment, and intuitive judgment.

Inactive Publication Date: 2016-06-22
浙江中迪生物科技有限公司
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Problems solved by technology

[0004] At present, the most common detection method for gene polymorphism is sequencing method, which is suitable for high-throughput multi-site detection, but it is time-consuming and low-sensitivity, so it is not suitable for rapid clinical detection; high-resolution melting curve method is not suitable for equipment The requirements are quite special, and it needs to be installed with high-resolution software and temperature-sensitive machines to be used, and there are certain difficulties in its clinical promotion

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  • SNP detecting kit for methionine-folic acid metabolism key enzymes MTHFR and use method thereof
  • SNP detecting kit for methionine-folic acid metabolism key enzymes MTHFR and use method thereof
  • SNP detecting kit for methionine-folic acid metabolism key enzymes MTHFR and use method thereof

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Embodiment approach

[0069] As an improved specific embodiment, the volume ratio of the primer and the probe is 1:1.

[0070] As an improved specific embodiment, the amounts of the forward primer and the reverse primer are the same.

[0071] As an improved specific embodiment, the MTHFRC677T wild-type probe and the MTHFRC677T mutant probe are used in the same amount.

[0072] As an improved specific embodiment, the MTHFRA1298C wild-type probe and the MTHFRA1298C mutant probe are used in the same amount.

[0073] The method for using the kit comprises the following steps:

[0074] 1) Extract the genomic DNA of the person to be tested;

[0075] 2) Each PCR system includes volume parts: 10-25 parts of master mix, 1 part of DNA template, 1-2 parts of primers, and 1-2 parts of probe;

[0076] 3) Put the configured PCR system into a fluorescent quantitative PCR instrument for Real-time PCR detection; the reaction conditions are:

[0077] Pre-denaturation at 95°C for 10 minutes; cycle stage, 95°C for...

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Abstract

The invention discloses an SNP detecting kit for methionine-folic acid metabolism key enzymes MTHFR and a use method thereof. Corresponding specific primers and fluorescence probes for detecting MTHFR C677T polymorphism and MTHFR A1298C polymorphism are added, a real-time fluorescence quantification PCR system is formed, and through a real-time PCR, technicists in the field determine and detect the genotype of a corresponding MTHFR gene locus through data collected by a fluorogenic quantitative PCR amplifier. The advantages of being fast and sensitive, facilitating judgment and the like are achieved, and the SNP detecting kit and the use method thereof are suitable for clinical detection and popularization.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a SNP detection kit for the key enzyme of methionine-folate metabolism MTHFR and a use method thereof. Background technique [0002] The full name of MTHFR is methylenetetrahydrofolate reductase, which is a key enzyme in the metabolism of methionine-folate, which can reduce 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, thereby As an indirect donor of methyl group, it participates in the synthesis of purine and pyrimidine in the body and the methylation of DNA, RNA, and protein, while maintaining the normal level of homocysteine ​​in the body. [0003] A large number of studies have confirmed that the mother's MTHFRC677T and MTHFRA1298C mutations are genetic risk factors for children with neural tube defects. MTHFRC677T and MTHFRA1298C reduce the enzyme activity of MTHFR, causing plasma hyperhomocysteinemia. Hyperhomocysteinemia during pregnancy can lead to a hyper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2561/113C12Q2563/107
Inventor 邢洋苏彩环林芳珍
Owner 浙江中迪生物科技有限公司
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