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Kit for genetic detection of gout

A kit and technology for gout, applied in the fields of molecular biology and medicine, can solve the problems of folic acid metabolism disorder, reduced activity, and reduced heat tolerance

Inactive Publication Date: 2009-12-23
XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

C677T is located in the catalytic region of the MTHFR gene, and this mutation directly affects the activity and heat resistance of the MTHFR protein, manifested in varying degrees of reduced enzyme activity accompanied by reduced heat resistance of the enzyme
When the risk genotype is carried, it directly affects the activity and heat resistance of the MTHFR gene, manifested in varying degrees of reduced activity of 5,10-methylenetetrahydrofolate reductase accompanied by reduced heat resistance of the enzyme, and further triggers Cause folic acid metabolism disorder, affect the synthesis of purine, cause the increase of uric acid, and finally increase the risk of gout

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Use of detection kits

[0047] Step 1: Extraction of DNA template

[0048] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0049] Step 2: PCR amplification reaction

[0050] Use the PCR reaction set included in the detection kit.

[0051] The reaction system has a total volume of 25 μl, including 1 μl of DNA template with a concentration of 12.5ng / μl, 2.5 μl of 10XPCR reaction buffer, 0.2 μl of 25mM dNTP mixture, 1.5 μl of 25mM MgCl2 solution, 0.125 μl of 5units / μl Taq DNA polymerase, 20 μM 0.25 μl of each primer for each specific primer pair, and 19.175 μl of deionized water.

[0052] The reaction was carried out on the ABI2720 type PCR amplification instrument. The reaction conditions were 94°C, 12 minutes, and 30 cycles of 94°C, 30 seconds, 60°C, 30 seconds, 72°C, 30 seconds, and then 72°C, 10 minutes .

[0053] Step 3: PCR Product Purification

[0054] Use the PCR product purification kit in the detection kit. The reac...

Embodiment 2

[0068] Example 2. Application of detection kits for individual gout genetic susceptibility testing services

[0069] Step 1: DNA Extraction

[0070] 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.

[0071] Step 2: Genotyping Assays

[0072] Use the kit provided by the present invention to carry out DNA sequencing detection on the rs429358 SNP site and the rs7412 SNP site on the APOE gene of the subject's genomic DNA, and perform fluorescence quantitative PCR detection on the C677T SNP site on the MTHFR gene to determine this Genotypes of the 3 SNP loci.

[0073] Step 3: Analysis of Individual Genetic Susceptibility to Gout

[0074] Through the analysis of the SNP genotype of the tested subjects, an analysis report of the individual's genetic susceptibility to gout is issued. The report details the le...

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Abstract

The invention discloses a kit for genetic detection of gout. The kit comprises a specific primer pair and a specific probe pair which are used for detecting the mononucleotide polymorphism loci genotype of an apolipoprotein E gene (APOE), and a 5,10-methylenetetrahydrofolate reductase gene (MTHFR), a fluorescent quantitative PCR general component, a PCR reaction component and the like. The kit can evaluate the genetic risk of individuals suffered from the gout by detecting the polymorphism loci genotype of the genes closely related to the genetic risk of the gout at the same time.

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 the genetic susceptibility of individual gout, by simultaneously detecting the apolipoprotein E gene (APOE), 5,10- Single nucleotide polymorphism locus genotypes on the methylenetetrahydrofolate reductase gene (MTHFR) were used to assess individual genetic susceptibility to gout. Background technique [0002] Gout is a disease caused by chronic purine metabolism disorder in the body. It is mainly due to the increase of uric acid production and / or the decrease of uric acid excretion, and the deposition of uric acid in the body, resulting in pathophysiological changes. Manifested as hyperuricemia, recurrent gouty acute arthritis, tophi, uric acid nephropathy, often accompanied by urinary tract stones, severe joint deformities and dysfunction. [0003] However, different people have different genetic cond...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 冯哲民邹祖烨
Owner XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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