Human MTHFR (Methylene Tetrahydrofolate Reductase) and/or MTRR (Methylenetetrahydrofolate Reductase) gene polymorphism investigation kit

Abstract

The invention discloses a kit for investigating polymorphism of an MTHFR (Methylene Tetrahydrofolate Reductase) and / or MTRR (Methylenetetrahydrofolate Reductase) gene on the basis of a Taqman-MGB probe. The kit comprises a primer group and a probe, wherein the primer group is at least one selected from the following three groups: primers of SEQ ID NO.1 and SEQ ID NO.4 for an MTHFR gene at a C677Tsite, primers of SEQ ID NO.7 and SEQ ID NO.10 for an MTRR gene at an A1298C site, primers of SEQ ID NO.13 and SEQ ID NO.14 for an MTRR gene at an A66G site, primers of SEQ ID NO.22 and SEQ ID NO.19, primers of SEQ ID NO.29 and SEQ ID NO.15, and primers of SEQ ID NO.33 and SEQ ID NO.30. The kit disclosed by the invention has the advantages that three mutation sites can be detected simultaneously, high sensitivity is achieved, plasma as low as 10copies can be accurately detected, and oral cavity swabs which are too long in preservation time or relatively low in concentration can be still accurately detected.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a human MTHFR and MTRR gene polymorphism detection kit. Background technique [0002] Enzymes closely related to folate metabolism are 5,10-methylenetetrahydrofolate reductase (MTHFR) and methionine synthase reductase (MTRR). Genetic variations such as MTHFR and MTRR (mainly polymorphisms of C677T and A1298C of the MTHFR gene and A66G of the MTRR gene) can inhibit the conversion of homocysteine ​​to methionine, resulting in hypofolate and hyperhomocysteinemia, thereby increasing the risk of birth defects in newborns or the risk of spontaneous abortion. [0003] Folate deficiency is the main cause of birth defects in newborns. There are two reasons for the lack of folic acid in the body: one is insufficient folic acid intake, and the other is the low ability of the body to utilize folic acid (folic acid metabolic pathway disorder) due to ge...

AI Technical Summary

Problems solved by technology

This method can only detect mutations at enzyme cleavage sites, is time-consuming and laborious, has a long test cycle, and there is also the risk of false positives caused by PCR product contamination

[0007] (3) Chip method: PCR products need subsequent processing, cumbersome operation, expensive, and prone to false positives

[0009] ①HRM method: high-resolution melting curve analysis technology, which is to add DNA double-strand binding dye to the PCR system, and type the sample according to the melting curve. Because the fluorescent signal comes from the dye, the specificity is not high, and the price of the instrument is high. Popularity is limited

[0012] The Taqman probe method is also widely used in biotechnology at present. However, as the above-mentioned various detection methods have their advantages and disadvantages, as for the Taqman-MGB probe, there may only be 7-8 bp on both sides of the SNP site. , the mismatch of one base is enough to cause the annealing failure of the probe, and if the MGB probe is not well designed, the resolution will be similar to that of ordinary Taqman probes, but MGB probes are much more expensive than ordinary Taqman probes, so it is necessary to To truly realize the applicability of detection products based on Taqman-MGB probes, especially when it involves the parallel detection of more than one gene, creative design of the entire detection platform is required

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