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Kit for detecting individual folate metabolism disorder

A technology of folic acid metabolism and kit, which is applied in the fields of molecular biology and medicine, and can solve problems such as the reduction of enzyme activity

Inactive Publication Date: 2014-03-26
浙江爱易生物医学科技有限公司
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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

[0049] Example 1. Use of detection kits

[0050] Step 1: Extraction of DNA template

[0051] Vacuum blood collection tubes are used to collect human peripheral blood, and the genomic DNA in the blood is extracted.

[0052] Step 2: If you want to make N tubes, in a 1.5ml or larger volume tube, add ddH 2 For each reagent from O to SYBR, the volume is: N x single volume.

[0053] Operate on ice to maintain the activity of the reagents.

[0054] Because of the SYBR fluorescent dye, avoid moderate and strong light exposure.

[0055] Under the premise of accurate pipetting, the operating time should be shortened as much as possible to reduce non-specific amplification.

[0056] Shake well to even out ingredients.

[0057] Step 3: Add the template DNA and the mixed solution prepared in step 2 into the PCR tube, shake and centrifuge.

[0058] Step 4: Real-time quantitative PCR reaction

[0059] Use a real-time quantitative PCR detection kit that can detect genes related to foli...

Embodiment 2

[0077] Example 2. Services for detection of folic acid disorders in women of childbearing age

[0078] Step 1: DNA Extraction

[0079] The laboratory physicians of the hospital collect peripheral blood from the subjects, use vacuum blood collection tubes to collect peripheral blood, and extract genomic DNA from it.

[0080] Step 2: Genotyping Assays

[0081] Use the kit provided by the present invention to carry out real-time quantitative PCR detection respectively to the SNP site No. rs7216389 on the ORMDL3 gene of the subject's genomic DNA and the SNP site No. rs25681 on the C5 gene to determine the genes of these two SNPs sites type.

[0082] Step 3: Analysis of Individual Folate Metabolism Capacity

[0083] Through the analysis of the SNPs genotypes of the tested subjects, an analysis report of the individual folic acid metabolism ability is issued. The report details the subject's folic acid metabolism ability, and the physician explains and interprets the analysis re...

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PUM

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Abstract

The invention discloses a kit for detecting an individual folate metabolism disorder. The kit comprises a specific primer pair and SYBRGreenI fluorochrome for simultaneously detecting rs1801133# SNP site on a 5,10-methylene tetrahydrofolate reductase gene, rs1801131# SNP on a 5,10-methylene tetrahydrofolate reductase gene and rs1801394# SNP site on a methionine synthase reductase gene, a conventional component for real-time quantitative polymerse chain reaction (PCR) detection. The kit disclosed by the invention evaluates the metabolic capability of individual folate by simultaneously detecting single nucleotide polymorphism site genotype of the 5,10-methylene tetrahydrofolate reductase gene and the methionine synthase reductase gene of folate metabolism.

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 detecting individual folic acid metabolism disorders, through simultaneous detection of 5,10-methylenetetrahydrofolate reductase which is closely related to folic acid metabolism ability Gene (MTHFR), methionine synthase reductase gene (MTRR) single nucleotide polymorphism site genotype to assess individual folate metabolism capacity. Background technique [0002] Folic acid is an essential substance when the human body utilizes sugar and amino acids, and is an essential substance for the growth and reproduction of body cells. Folic acid acts in the form of tetrahydrofolate in the body, and tetrahydrofolate participates in the synthesis and transformation of purine nucleic acid and pyrimidine nucleotide in vivo. Folic acid plays an important role in the manufacture of nucleic acids (ribonucleic acid, deoxyribonuc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 杨玉华
Owner 浙江爱易生物医学科技有限公司
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