MTRR (5-methyltetrahydrofolate-homocysteine methyltransferase reductase) gene mutation detection specific primers and liquid chip

Abstract

The invention discloses an MTRR (5-methyltetrahydrofolate-homocysteine methyltransferase reductase) gene mutation detection liquid chip and specific primers. The liquid chip mainly comprises each ASPE (allele specific primer extension) primer formed by tag sequences at 5' terminal and specific primer sequences aiming at target gene mutation sites at 3' terminal, microspheres coated by anti-tag sequences as well as amplification primers, wherein the specific primer sequences are SEQ ID NO.13 and SEQ ID NO.14 aiming at G28905A sites, SEQ ID NO.15 and SEQ ID NO.16 aiming at C1783T sites, SEQ ID NO.17 and SEQ ID NO.18 aiming at A66G sites, SEQ ID NO.19 and SEQ ID NO.20 aiming at A1049G sites, SEQ ID NO.21 and SEQ ID NO.22 aiming at C1243T sites and / or SEQ ID NO.23 and SEQ ID NO.24 aiming at C1349G sites. The liquid chip has the advantages that the coincidence rate of the detection results of the detection liquid chip provided by the invention and a sequencing method is as high as 100%; and parallel detection of wild types and mutant types of a plurality of mutation sites is achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for MTRR gene mutation detection and a liquid phase chip. Background technique [0002] Methionine synthase reductase (5-methyltetrahydrofolate-homocysteine ​​methyltransferase reductase, MTRR), the MTRR gene is located on the short arm of human autosome 5 (5P15.2-15.3), the gene contains 15 exons and 14 introns. The A66G mutation of the MTRR gene is the most important and most studied mutation site at present. The polymorphism of the A66G site is related to the occurrence of spina bifida, which will affect the concentration of homocysteine ​​and folic acid metabolism. Whether there is the possibility of neural tube defects in future offspring, and predict the effect of angiotensin-converting enzyme inhibitor drugs. In addition, the G28905A, C1783T, A1049G, C1243T and C1349G site mutations of the MTRR gene are also relate...

AI Technical Summary

Problems solved by technology

[0003] At present, MTRR gene mutation detection methods mainly include: PCR-RFLP, direct sequencing method and fluorescent quantitative PCR technology. PCR-RFLP method is based on the change of restriction enzyme recognition site caused by gene mutation, such as site loss or generation New site, a specific fragment is amplified by PCR, and then the amplified product is digested with restriction endonuclease, and the size of the fragment is observed by electrophoresis. Genotype, but this method cannot be used for the detection of gene mutations that do not generate new restriction sites

However, other PCR-based detection technologies, such as direct sequencing and fluorescent quantitative PCR technology, have the disadvantages of low sensitivity, easy contamination of samples, and high false positive rate.

Again, the above methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

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