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Primer probe combination for detecting polymorphism of folate metabolism related genes and application of primer probe combination

A technology for gene polymorphism and folic acid metabolism, which is applied in the field of primer-probe combinations for detecting gene polymorphisms related to folic acid metabolism, can solve the problems of restricting the popularization and use of methods, high requirements for technicians, complicated operations, etc. Human operation error, saving detection time, simple and convenient operation

Pending Publication Date: 2022-04-15
SHANGHAI BIOTECAN PHARMA +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method requires purification of samples and sequencing, cumbersome operation, high cost, and needs to be used with professional instruments, which requires high technical personnel, which limits the promotion and use of related methods

Method used

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  • Primer probe combination for detecting polymorphism of folate metabolism related genes and application of primer probe combination
  • Primer probe combination for detecting polymorphism of folate metabolism related genes and application of primer probe combination
  • Primer probe combination for detecting polymorphism of folate metabolism related genes and application of primer probe combination

Examples

Experimental program
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Effect test

Embodiment 1

[0087] This example provides a combination of primers and probes for detecting polymorphisms of genes related to folic acid metabolism. The combination of primers and probes for detecting polymorphisms of genes related to folic acid metabolism specifically amplifies and detects MTHFR gene C677T, MTHFR gene A1298C and The polymorphism of MTRR gene A66G;

[0088] The upstream primer of the specific primer pair for amplifying the MTHFR gene C677T is the nucleotide sequence shown in SEQ ID No.1, and the downstream primer is the nucleotide sequence shown in SEQ ID No.2 to detect the MTHFR gene The probe of C677T is the nucleotide sequence shown in SEQ ID No.3;

[0089] The upstream primer of the specific primer pair for amplifying the MTHFR gene A1298C is the nucleotide sequence shown in SEQ ID No.4, and the downstream primer is the nucleotide sequence shown in SEQ ID No.5 to detect the MTHFR gene The probe of A1298C is the nucleotide sequence shown in SEQ ID No.6;

[0090] The u...

Embodiment 2

[0105] This embodiment provides a kit for detecting gene polymorphisms related to folic acid metabolism. Needle combinations, where:

[0106] The final concentration of the upstream primer of the specific primer pair for amplifying the MTHFR gene C677T is 0.01 μmol / L, the final concentration of the downstream primer is 0.2 μmol / L, and the final concentration of the probe for detecting the MTHFR gene C677T is 0.05 μmol / L;

[0107] The final concentration of the upstream primer of the specific primer pair for amplifying the MTHFR gene A1298C is 0.06 μmol / L, the final concentration of the downstream primer is 0.006 μmol / L, and the final concentration of the probe for detecting the MTHFR gene A1298C is 0.01 μmol / L;

[0108] The final concentration of the upstream primer of the specific primer pair for amplifying the MTRR gene A66G is 0.3 μmol / L, the final concentration of the downstream primer is 0.03 μmol / L, and the final concentration of the probe for detecting the MTRR gene...

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Abstract

The invention provides a primer probe combination for detecting the polymorphism of a folate metabolism related gene and application of the primer probe combination. The primer probe combination for detecting the polymorphism of the folate metabolism related gene specifically amplifies and detects the polymorphism conditions of an MTHFR (Methylene Tetrahydrofolate Reductase) gene C677T, an MTHFR gene A1298C and an MTRR (Methylene Tetrahydrofolate Reductase) gene A66G; an upstream primer of a specific primer pair for amplifying the MTHFR gene C677T comprises a nucleotide sequence as shown in SEQ ID No. 1, a downstream primer of the specific primer pair for amplifying the MTHFR gene C677T comprises a nucleotide sequence as shown in SEQ ID No. 2, and a probe for detecting the MTHFR gene C677T comprises a nucleotide sequence as shown in SEQ ID No. 3. The invention also provides a kit for detecting the polymorphism of the folate metabolism related gene. The primer probe combination is high in sensitivity, good in specificity, high in detection efficiency, easy to operate and wide in application prospect.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection, and in particular relates to a combination of primers and probes for detecting gene polymorphisms related to folic acid metabolism and its application. Background technique [0002] Folic acid, also known as vitamin B9, plays an important role in cell growth and the synthesis of nucleic acids, amino acids, and proteins. Folic acid deficiency can lead to hypofolate and hyperhomocysteinemia, thereby increasing the risk of birth defects and cardiovascular diseases in newborns. MTHFR and MTRR are key enzymes in the process of folic acid metabolism. Mutations in the MTHFR gene will affect the enzyme activity and thermal stability of MTHFR, thereby affecting the metabolism of folic acid and methionine. Mutations in the MTRR gene will cause changes in the activity of the MTRR enzyme, leading to folate deficiency or elevated homocysteine ​​levels in the body. [0003] Therefore, it is of...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
Inventor 王博伟汪梦竹吴煜周凯月何顺清林灵
Owner SHANGHAI BIOTECAN PHARMA
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