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Kit for genetic detection of breast cancer

A kit and breast cancer technology, applied in the field of breast cancer genetic detection kits, can solve problems such as inability to stabilize genetic information of the genome, influence of BRCA2 function, and increased risk of breast cancer

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

When carrying this mutant genotype (-- / -- or CT / --), the function of BRCA2 is affected, and the stability of the genetic information of the genome cannot be maintained through DNA repair, and the risk of breast cancer increases

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Use of detection kits

[0033] Step 1: Extraction of DNA template

[0034] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0035] Step 2: Real-time quantitative PCR reaction

[0036] Use the fluorescent quantitative PCR kit in the detection kit to perform 8 independent fluorescent quantitative PCR reactions, each reaction system has a total volume of 10 μl, and contains 2 μl of DNA template with a concentration of 20 ng / μl, and 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.

[0037] 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 sec...

Embodiment 2

[0041] Example 2. Application of detection kits for individual breast cancer genetic susceptibility testing services

[0042] Step 1: DNA Extraction

[0043] 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.

[0044] Step 2: Genotyping Assays

[0045] Using the kit provided by the present invention, the Pvu II SNP site on the ER-α gene of the subject's genomic DNA, the XbaI SNP site on the ER-α gene, the Val158Met SNP site on the COMT gene, and the Ile105Val SNP site on the GSTP1 gene The 1100delATSNP site on the BRCA1 gene, the 589delCT SNP site on the BRCA1 gene, the T5319A SNP site on the BRCA1 gene, and the 5950delCT SNP site on the BRCA2 gene were detected by fluorescent quantitative PCR to determine the genotypes of these 8 SNP sites.

[0046] Step 3: Analysis of Individual Genetic Susceptibility t...

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PUM

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Abstract

The invention discloses a kit for genetic detection of breast cancer. The kit comprises a specific primer pair and a specific probe pair which are used for detecting the mononucleotide polymorphism loci genotype of an estrogen receptor alpha gene (ER-alpha), a catechol-o-methyltransferas gene (COMT), a glutathione-s-transferase P1 gene (GSTP1), a human breast cancer susceptibility gene 1 (BRCA1), and a human breast cancer susceptibility gene 2(BRCA2), a fluorescent quantitative PCR general component and the like. The kit can evaluate the genetic risk of individuals suffered from the breast cancer by detecting the polymorphism loci genotype of the genes closely related to the genetic risk of the breast 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 breast cancer, by simultaneously detecting the estrogen receptor α gene (ER-α) closely related to breast cancer , catechol oxygen methyltransferase gene (COMT), glutathione sulfur transferase P1 gene (GSTP1), human breast cancer susceptibility gene 1 (BRCA1), human breast cancer susceptibility gene 2 (BRCA2) Single nucleotide polymorphism locus genotypes to assess individual genetic susceptibility to breast cancer. Background technique [0002] Breast cancer is a disease in which ductal epithelial cells of the mammary gland lose their normal characteristics and proliferate abnormally under the action of various internal and external carcinogenic factors, so that they exceed the limit of self-repair and become cancerous. The main clinical manifestation is breast lu...

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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