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Kit for genetic detection of colorectal cancer

A kit and technology for colorectal cancer, applied in the field of colorectal cancer genetic detection kits, can solve problems such as increased risk of colorectal cancer, changes in protein space conformation, and the impact of gene damage repair function

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 reduction of the amount of 5,10-methylenetetrahydrofolate is not conducive to the conversion of dUMP into dTMP, which may lead to the incorporation of base U in DNA synthesis (impeding DNA synthesis)
When carrying the risk genotype (AA), the spatial conformation of hMLH1 protein changes, the gene damage repair function is affected, and the risk of colorectal cancer increases

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1. Use of detection kits

[0039] Step 1: Extraction of DNA template

[0040] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0041] Step 2: Electrophoretic detection and typing

[0042] Using the electrophoresis detection kit in the detection kit, the reaction system has a total volume of 25 μl, including 1 μl of DNA template with a concentration of 12.5 ng / μl, 2.5 μl of 10X PCR reaction buffer, 0.2 μl of 25mM dNTP mixture, and 1.5 μl of 25mM MgCl2 solution , 0.125 μl of 5 units / μl Taq DNA polymerase, 0.25 μl of each primer for 20 μM electrophoresis detection primer pair, and 18.675 μl of deionized water.

[0043] The reaction was carried out on a PCR amplification instrument, and the reaction conditions were 94° C. for 12 minutes, and 30 cycles of 94° C. for 30 seconds, 60° C. for 30 seconds, 72° C. for 30 seconds, and then 72° C. for 10 minutes.

[0044] Then use electrophoresis technology to perform agarose gel electrophoresis...

Embodiment 2

[0052] Example 2. Application of detection kits for individual colorectal cancer genetic susceptibility testing services

[0053] Step 1: DNA Extraction

[0054] 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.

[0055] Step 2: Genotyping Assays

[0056] Use the kit provided by the invention to carry out PCR amplification and electrophoresis detection on the GSTT1 gene and the SNP site on the GSTM1 gene of the genomic DNA of the subject to be tested, and perform PCR amplification and electrophoresis detection on the C677T SNP site on the MTHFR gene and the Rsa I SNP site on the CYP2E1 gene. , T1151A SNP sites on the hMLH1 gene were detected by fluorescent quantitative PCR to determine the genotypes of these 5 SNP sites.

[0057] Step 3: Analysis of Individual Genetic Susceptibility to Colorectal Cancer ...

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PUM

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Abstract

The invention discloses a kit for genetic detection of colorectal cancer. The kit comprises a specific primer pair and a specific probe pair which are used for detecting the mononucleotide polymorphism loci genotype of a 5,10-methylenetetrahydrofolate reductase gene (MTHFR), a glutathione-s-transferase T1 gene (GSTT1), a glutathione-s-transferase M1 gene (GSTM1), a cytochrome P450 2E1 gene (CYP2E1), and a mismatching repair enzyme gene (hMLH1), 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 colorectal cancer by detecting the polymorphism loci genotype of the genes closely related to the genetic risk of the colorectal cancer 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 colorectal cancer. enzyme gene (MTHFR), glutathione sulfur transferase T1 gene (GSTT1), glutathione sulfur transferase M1 gene (GSTM1), cytochrome P450 2E1 gene (CYP2E1), mismatch repair gene (hMLH1) Single nucleotide polymorphism locus genotypes to assess individual genetic susceptibility to colorectal cancer. Background technique [0002] Colorectal cancer is the general term for colon cancer and rectal cancer. It refers to the canceration of the large intestine mucosal epithelium under the action of various carcinogenic factors such as environment or genetics. It is one of the common malignant tumors and has a high mortality rate. As cancer cells multiply, the smooth bowel wall becomes rough, swollen and hard, and eventually ulcers develop. The ulcerated area...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 冯哲民邹祖烨
Owner XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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