MTRR gene polymorphism detection probe, primer, kit and detection method

Abstract

The invention relates to an MTRR gene polymorphism detection probe, a primer, a kit and a detection method. The MMRR gene polymorphism detection probe comprises a probe T1 used for detecting the A66Gsite of the MTRR gene, and the nucleotide sequence of the probe is shown in SEQ ID No. 1. The MMRR gene polymorphism detection probe, the primer and the kit have the advantages of non-invasive sampling, no DNA extraction, low cost, high efficiency, accuracy, simple and convenient operation, clear result interpretation and easy clinical popularization.

Description

technical field [0001] The invention relates to gene detection technology, in particular to a detection probe, primer, kit and detection method for MTRR gene polymorphism. Background technique [0002] Folic acid belongs to B vitamins. It is an essential element for the synthesis of nucleic acid, a substance necessary for cell growth and tissue repair, and an indispensable nutrient during embryonic development. Folic acid deficiency is one of the main causes of birth defects. Folic acid deficiency Mostly caused by genetic defects, the body's metabolism of folic acid is impaired. [0003] Folate plays an important role in the homocysteine ​​metabolic pathway. Folic acid acts in the form of tetrahydrofolate in the body. If folic acid is deficient and the synthesis of N5-methyltetrahydrofolate is insufficient, the conversion of homocysteine ​​to methionine will be impaired, and high concentrations of homocysteine ​​will occur. Amino acid can damage the endothelial cells of bl...

AI Technical Summary

Problems solved by technology

However, this method has the defects of long time-consuming, high risk of contamination and poor specificity; (6) Mutation amplification retardation PCR (AMRS): also known as allele-specific amplification method, using the 3` terminal position of PCR primers The principle that the base must be complementary to its template DNA can be effectively amplified. Design allele-specific amplification primers. Only when the primer 3` base is paired with the template can the amplification band appear, thereby detecting mutations

This method needs to strictly control the reaction conditions, and the multiplex PCR amplification is more difficult, thus limiting the clinical application

[0008] In the prior art, the Chinese patent application with publication number 106957903A discloses a genotyping kit for detecting polymorphic sites of key enzymes in folic acid metabolism and its detection method, which uses the fluorescence melting curve method to detect gene sites including MTHFR:C677T, MTHFR:A1298C, and MTRR:A66G; specifically, it is designed with 3 probes to detect 3 mutation sites respectively, the number of probes is large, the cost is high, multi-channel fluorescence detection is required, and the TM value Affected by the detection sequence of the probe, the design and detection effect are not good

[0009] The Chinese patent application with publication number 106701987A discloses a PCR amplification system and detection kit for genotyping three SNP loci related to human folic acid metabolism, which uses the AMRS-PCR method to detect gene loci including MTHFR:C677T , MTHFR:A1298C and MTRR:A66G; the specificity of the above-mentioned detection scheme is poor, the requirement for the ion level in the system is high, and the DNA purity is greatly affected, so it is not suitable for clinical application

[0010] The Chinese patent application with the publication number 107254520A discloses a detection method for genes related to folic acid metabolism, which adopts Sanger sequencing method, and detects gene loci including MTHFR:C677T, MTHFR:A1298C and MTRR:A66G; the above-mentioned detection scheme is cumbersome to operate, and Pollution from opening the cap

[0011] The Chinese patent application with publication number 104131087A discloses a kit for genotyping folic acid metabolism genes, which uses the reverse dot hybridization method to detect gene loci including MTHFR:C677T, MTHFR:A1298C and MTRR:A66G; the above The detection scheme is cumbersome, time-consuming, and has poor specificity

[0012] The Chinese patent application with the publication number 107058530A discloses a kit for direct detection of folic acid drug-sensitive genes in blood and its use method, which uses fluorescent PCR method, and the detection gene loci include MTHFR:C677T and MTHFR:A1298C; the above The detection scheme requires multiple probes and requires many channels, and there are few suitable fluorescent PCR equipment, which is not suitable for clinical promotion

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