MTHFR, MTRR and RFC1 gene polymorphism detection primer combination and kit and application of MTHFR, MTRR and RFC1 gene polymorphism detection kit

Abstract

The invention discloses an MTHFR, MTRR and RFC1 gene polymorphism detection primer combination and kit and application of the MTHFR, MTRR and RFC1 gene polymorphism detection kit. The detection kit comprises ultrapure water, an X solution, a 10*PCR buffering solution, a PCR primer, a 25 mM magnesium chloride solution, DNA polymerase and a positive control product, wherein the PCR primer comprises a forward and reverse amplification primer, a DNA internal reference forward and reverse amplification primer and a reaction internal reference forward and reverse amplification primer of different gene types, the three primers are located at four SNP sites on a gene related to folic acid metabolism, and the gene sequence of the PCR primer is shown in SEQ ID NO.1-NO.20. The detection kit has the advantages of being high in specificity, accuracy, flux and reliability, low in cost and free of a false-negative result.

Description

technical field [0001] The invention relates to a multiple gene detection kit and its application, in particular to a MTHFR, MTRR and RFC1 gene polymorphism detection kit and its application. Background technique [0002] Folic acid belongs to the B vitamins, and it is an essential element for the synthesis of nucleic acid, an essential substance for cell growth and tissue repair, and an indispensable nutrient for embryonic development. Numerous studies in recent years have confirmed that folic acid deficiency is the main cause of birth defects. In addition to preventing fetal neural tube defects, the clinical function of folic acid can also reduce the incidence of pregnancy-induced hypertension, spontaneous abortion, intrauterine growth retardation, premature delivery, and low birth weight in newborns. [0003] And China is one of the countries with high incidence of birth defects in the world, and the number of children with birth defects every year accounts for about 20%...

AI Technical Summary

Problems solved by technology

Its disadvantages are (1) low throughput: it is not suitable for the detection of multiple SNP sites; it is difficult to set internal control genes

(2) High cost: the cost of probe labeling is high; if you need to obtain all relevant SNP information, you need to conduct multiple detection tests, and the superimposed cost is more expensive

Disadvantages: 1) The hybridization kinetics of different SNP sites are different, and the conditions are difficult to control when performing simultaneous detection of multiple sites; 2) The technology is expensive and complicated: each sample requires a chip, and the cost is more than ￥1000 / sample , which is not conducive to large-scale promotion; the synthesis and fixation of probes are more complicated, especially making high-density probe arrays, which is the main speed-limiting step; 3) poor repeatability, low accuracy, and false positive and false negative results are prone to occur; 4) Low sensitivity: the chip method requires a large amount of nucleic acid, and generally requires multiple PCR amplification first. Due to the large number of primers, it is easy to produce dimers, hairpin structures, or the purpose of amplification due to different Tm values. Different fragment efficiencies affect the sensitivity of detection; 5) Due to the variety of chips, it is difficult to formulate a unified quality control standard

[0015] At present, there are no relevant reports on kits and detection methods for multiplex SNP detection based on multiplex PCR and CE separation to guide folic acid medication at home and abroad

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