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Kit for quantitative guidance of folic acid supplement dosage before and during pregnancy

A kit and folic acid technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve problems such as decreased enzyme activity

Inactive Publication Date: 2010-06-30
HAINAN ZHUJIAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

MTRR 66A→G polymorphism, resulting in replacement of methionine with isoleucine, resulting in reduced enzyme activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Use of detection kits

[0029] Step 1: Extraction of DNA template

[0030] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0031] Step 2: Real-time quantitative PCR reaction

[0032] Use the detection kit to carry out 3 independent fluorescent quantitative PCR reactions. The total volume of each reaction is 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10X fluorescent quantitative PCR reaction buffer, and 0.1 μl of 25 mM dNTP mixture. solution, 0.6μl 25mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA polymerase, 0.225 μl each of 20 μM sense primer and antisense primer, 0.25 μl each of 10 μM fluorescent probe with VIC and fluorescent probe with FAM, and 5.325 μl of deionized water.

[0033] The reaction was carried out on a PCR amplification instrument, and the reaction conditions were 50° C. for 2 minutes, 95° C. for 10 minutes, and 60 cycles of 95° C. for 30 seconds and 60° C. for 1 minute....

Embodiment 2

[0037]Example 2. Application of detection kits for quantitative guidance of folic acid supplements

[0038] Step 1: DNA Extraction

[0039] The physicians in the laboratory department of the hospital instructed the subjects to use oral swabs to sample oral epithelial cells, and the silica gel adsorption method was used to extract DNA from oral epithelial cells.

[0040] Step 2: Genotyping Assays

[0041] Using the kit provided by the invention, the rs1801133 and rs1801131 SNP sites on the 5,10-methylenetetrahydrofolate reductase gene and the rs1801394 methionine synthetase reductase gene on the genomic DNA of the subject were detected The SNP sites were detected by fluorescent quantitative PCR to determine the genotypes of the three SNP sites.

[0042] Step 3: Quantitative Guidelines for Folic Acid Supplements

[0043] Through the analysis of the SNP genotype of the tested person, an analysis report of the genetic ability of folic acid metabolism is issued. The report deta...

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PUM

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Abstract

The invention discloses a kit for quantitative guidance of folic acid supplement dosage before and during pregnancy and application thereof. The kit comprises a specific primer pair, a specific fluorescent probe pair, a convention assembly and the like, wherein the specific primer pair and a specific fluorescent probe pair can simultaneously detect three single nucleotide polymorphism genotypes on a 5, 10-methylene tetrahydrofolate reductase (MTHFR) gene and a methionine synthase reductase (MTRR) gene, and the convention assembly is used to for fluorescent quantitative PCR detection. The kit of the invention can be used for the quantitative guidance of the folic acid supplement dosage before and during the pregnancy.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, the present invention relates to a kit for quantitatively guiding the dosage of folic acid before pregnancy and during pregnancy. Folate reductase gene (MTHFR) and methionine synthase reductase gene (MTRR) single nucleotide polymorphism locus genotype to determine the folic acid supplement dosage suitable for different populations. Background technique [0002] Folic acid is an important nutrient needed by the body. It is a water-soluble B vitamin and is rich in green leafy vegetables, fruits and animal liver. Folic acid works in the human body in the form of tetrahydrofolate. Tetrahydrofolate is a carrier of one-carbon units, which can carry one-carbon units such as methyl, methylene, and formaldehyde, and participate in the synthesis of amino acids, purines, and pyrimidines in the body. Therefore, folic acid is an indispensable nutrient for cell divisio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 冯哲民邹祖烨
Owner HAINAN ZHUJIAN BIOTECH
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