Method for detecting polymorphism of folate metabolism-related genes

A technology for gene polymorphism and folic acid metabolism, which is applied in the field of detection of folic acid metabolism-related gene polymorphisms, can solve the problems of low efficiency and long cycle of detection of folic acid metabolism ability

Active Publication Date: 2019-08-23
BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the PCR-direct sequencing method takes a long time to detect folic acid metabolism in patients, which leads to low detection efficiency of folic acid metabolism in patients

Method used

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  • Method for detecting polymorphism of folate metabolism-related genes
  • Method for detecting polymorphism of folate metabolism-related genes
  • Method for detecting polymorphism of folate metabolism-related genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0167] In Example 1, a method for detecting gene polymorphisms related to folic acid metabolism may include:

[0168]Step C1: Pre-synthesize specific amplification primers and single-base extension primers for the C677T site, the A1298C site of the MTHFR gene, and the A66G site of the MTRR gene.

[0169] Step C2: extract the genomic DNA of the sample to be tested from the sample to be tested, and use the genomic DNA of the sample to be tested as a DNA template, wherein the concentration of the DNA template is 125 ng / μl.

[0170] Among them, the genomic DNA of the sample to be tested is extracted from the peripheral blood sample of the sample to be tested using a blood / cell / tissue genomic DNA extraction kit, and the concentration is measured by NP80-touch in the range of 5 to 1000 ng / μl, and to be The OD260 / OD280 ratio of the genomic DNA of the test sample was in the range of 1.6 to 2.2.

[0171] Step C3: Prepare 0.9 μl of distilled water, 25 mmol of MgCl 2 0.4 μl, 0.1 μl of ...

Embodiment 2

[0182] In embodiment 2, a method for detecting gene polymorphisms related to folic acid metabolism may include:

[0183] Step D1: pre-synthesizing specific amplification primers and single-base extension primers for the C677T site, the A1298C site of the MTHFR gene, and the A66G site of the MTRR gene.

[0184] Step D2: extract the genomic DNA of the test sample from the test sample, and use the test sample genomic DNA as a DNA template, wherein the concentration of the DNA template is 5 ng / μl.

[0185] Among them, the genomic DNA of the sample to be tested is extracted from the oral exfoliated cells of the sample to be tested using the Tiangen Oral Swab Genomic DNA Extraction Kit (DP322), and the concentration is measured by NP80-touch in the range of 5 to 1000 ng / μl Genomic DNA with OD260 / OD280 ratio between 1.6 and 2.2.

[0186] Step D3: Prepare 0.8 μl of distilled water, 25 mmol of MgCl 2 0.3 μl, 0.1 μl of 20 mmol deoxyribonucleoside triphosphate mixture, 0.9 μl of a mixt...

Embodiment 3

[0197] In embodiment 3, a method for detecting gene polymorphisms related to folic acid metabolism may include:

[0198] Step E1: pre-synthesizing specific amplification primers and single-base extension primers for the C677T site, the A1298C site of the MTHFR gene, and the A66G site of the MTRR gene.

[0199] Step E2: extract the genomic DNA of the sample to be tested from the sample to be tested, and use the genomic DNA of the sample to be tested as a DNA template, wherein the concentration of the DNA template is 119 ng / μl.

[0200] Among them, the genomic DNA of the sample to be tested is extracted from the tissue sample of the sample to be tested by using the blood / cell / tissue genomic DNA extraction kit (DP304), and the concentration is determined by NP80-touch within the range of 5-1000ng / μl , and the OD260 / OD280 ratio is within the range of 1.6 to 2.2.

[0201] Step E3: Prepare 1 μl of distilled water, 25 mmol of MgCl 2 0.5 μl, 0.2 μl of 20 mmol deoxyribonucleoside tri...

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Abstract

The invention provides a method for detecting polymorphism of folate metabolism-related genes. The method comprises the steps that specific amplification primers and single base extension primers of the C677T site and A1298C site of an MTHFR gene and the A66G site of an MTRR gene are synthesized in advance; to-be-detected sample genomic DNA is extracted from a to-be-detected sample to serve as a DNA template; a polymerase chain reaction PCR amplification system containing the specific amplification primers and the DNA template is prepared; a PCR apparatus is used for amplifying the PCR amplification system; digestion treatment is performed in sequence; a single base extension reaction system containing the single base extension primers is added into a reaction system for extension reactions in sequence, desalted purification treatment is performed in sequence, and a purified product is obtained; a detection instrument containing a mass spectrometer is used for detecting the purified product to determine the gene polymorphism of the C677T site and A1298C site of the MTHFR gene and the A66G site of the MTRR gene of the to-be-detected sample. The method for detecting the polymorphismof the folate metabolism-related genes can improve the detection efficiency of folate metabolism capacity in the body of a patient.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a method for detecting gene polymorphisms related to folic acid metabolism. Background technique [0002] Folic acid is an essential substance in the growth, division and reproduction of human cells. Studies have shown that the two appearances of folic acid in early embryonic development play an important role in genetic programming. Methylenetetrahydrofolate reductase (5,10-methylenetetrahydrofolate reductase, MTHFR) and methionine synthesis reductase (methioninesynthasereductase, MTRR) are the key enzymes of folic acid metabolism pathway. It has been found that there are multiple gene polymorphisms in the MTHFR and MTRR genes, and these gene polymorphisms affect the activity of the encoded enzymes, resulting in changes in serum folic acid. [0003] At present, when detecting the metabolic capacity of folic acid in a patient through the polymorphic sites of the MT...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6858
CPCC12Q1/6883C12Q1/6858C12Q2600/156C12Q2600/106C12Q2531/113C12Q2521/525C12Q2533/101C12Q2565/627Y02A50/30
Inventor 赵方圆佟洪梅智慧芳倪君君
Owner BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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