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Kit for genetic detection of diabetes

A kit and diabetes technology, applied in the fields of molecular biology and medicine, can solve the problems of weakening the ability of cellular immune response, decreasing the activity of vitamin D, weakening the regeneration and repair ability of pancreatic beta cells, etc.

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

The mutation of this gene will lead to the decline of vitamin D activity, weaken the ability of cellular immune response, weaken the regeneration and repair ability of islet β cells, reduce insulin secretion, and induce diabetes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1. Use of detection kits

[0028] Step 1: Extraction of DNA template

[0029] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0030] Step 2: Real-time quantitative PCR reaction

[0031] Use the fluorescent quantitative PCR kit in the detection kit to perform 4 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 10 X fluorescent quantitative PCR reaction buffer, 0.1μl 25mM dNTP mix, 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.

[0032] 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° ...

Embodiment 2

[0036] Example 2. Application of detection kits for individual diabetes genetic susceptibility testing services

[0037] Step 1: DNA Extraction

[0038] 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.

[0039] Step 2: Genotyping Assays

[0040] Use the kit provided by the invention to carry out the Gly15GlySNP site on the ADIPOQ gene of the subject's genomic DNA, the G-572C SNP site on the IL-6 gene, the PvuII SNP site on the LPL gene and the FokI SNP site on the VDR gene Real-time quantitative PCR detection was performed to determine the genotypes of the four SNP sites.

[0041] Step 3: Analysis of individual genetic susceptibility to diabetes

[0042] Through the analysis of the SNP genotype of the tested subjects, an analysis report sheet of the individual's genetic susceptibility to diabetes is is...

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Abstract

The invention discloses a kit for genetic detection of diabetes. The kit comprises a specific primer pair and a specific probe pair which are used for detecting the mononucleotide polymorphism loci genotype of an adiponection gene (ADIPOQ), an interleukin 6 gene (IL-6), a lipoprotein lipase gene (LPL) and a vitamin D receptor gene (VDR), a fluorescent quantitative PCR general component and the like. The kit can evaluate the genetic risk of individuals suffered from the diabetes by detecting the polymorphism loci genotype of the genes closely related to the genetic risk of the diabetes 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 diabetes. -6), lipoprotein lipase gene (LPL), vitamin D receptor gene (VDR) on the single nucleotide polymorphism site genotype to assess the individual genetic susceptibility to diabetes. Background technique [0002] Diabetes is a disease caused by a series of metabolic disorders of sugar, protein, fat, water and electrolytes due to insufficient or relatively insufficient insulin secretion and insulin resistance. Clinically, hyperglycemia is the main sign, and it usually manifests as "three excesses and one deficiency", that is, polyphagia, polyuria, polydipsia, and weight loss. In severe cases, acute metabolic disorders such as ketoacidosis can occur. [0003] Diabetes is not directly fatal, but prolonged illness can cause damage to multiple systems. Common ...

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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