Quick amplification detection kit of SNP (single nucleotide polymorphism) site of MTRR (5-methyltetrahydrofolate-homocysteinemethyltransferase reductase) and detection method

Abstract

The invention discloses a quick amplification detection kit of an SNP (single nucleotide polymorphism) site of MTRR (5-methyltetrahydrofolate-homocysteinemethyltransferase reductase) and a detection method. The kit comprises an efficient amplification group and a pyrophosphate sequencing group, wherein the efficient amplification group is used for linear exponent PCR (polymerase chain reaction) amplification of the SNP site of MTRR A66G; an amplification product is sequenced by the pyrophosphate sequencing group; a sequence of an amplification primer is shown as SEQ ID NOs (sequence identifiernumbers): 2 and 3 in a sequence table; and a sequence of an annealing primer is shown as SEQ ID NO: 4. A target segment linear exponent amplification method is adopted; the precisely designed amplification primer is high in amplification efficiency and low in mismatching rate; the amplification efficiency is greatly improved; single-stranded DNA (deoxyribonucleic acid) amplified directly can be directly used for pyrophosphate sequencing; the amplification product is not required to be subjected to labeling and double-chain separation; redundant experimental procedures are avoided; and the damage of a strongly basic reagent to an amplification segment is also reduced.

Description

technical field [0001] The invention relates to a rapid amplification detection kit and detection method of SNP sites of MTRR, belonging to the field of biotechnology, especially the field of biological sample detection. Background technique [0002] Methionine synthase reductase MTRR (5-methyltetrahydrofolate-homocysteinemethyltransferase reductase), the full name is encoding 5-methyltetrahydrofolate-homocysteine ​​methyltransferase reductase. Methionine is an essential amino acid for protein synthesis and one-carbon metabolism. Its synthesis is catalyzed by methionine synthase (encoded by the MTR gene), and methionine synthase is eventually oxidized due to the oxidation of the cofactor vitamin B12. Inactivate. The methionine synthase reductase encoded by MTRR can regenerate functionally active methionine synthase through reductive methylation. MTRR mutation is the main cause of folic acid / methyl vitamin deficiency type E, and it is also one of the main causes of abnormal...

AI Technical Summary

Problems solved by technology

[0004] The full length of MTRR is 32,021kb, with a total of 15 exons. It is located at the 5p15.3-p15.2 position of human chromosome 5, and its most important SNP site is A66G. The commonly used detection methods are post-amplification sequencing or gene chip Sequencing, but these detection methods have long detection time, cumbersome steps, large sample size and high detection cost, which greatly increase the financial burden of the testee, and the long-term result waiting also has a certain impact on the psychology of pregnant women and patients. Therefore, , based on years of research on pyrosequencing, the inventor has developed a rapid amplification detection kit and detection method for SNP sites of MTRR based on pyrosequencing

Images