Esophageal cancer genetic test kit

Abstract

The invention discloses a kit for testing the genetic risk of esophageal cancer, which comprises specific premier pairs for testing the single nucleotide polymorphic locus genotypes of a cytochrome P450 1A1 gene (CYP1A1), a cytochrome P450 2E1 gene (CYP2E1), a glutathione-S-transferase M1 gene (GSTM1), a 5,10-methylenetetrahydrofo late reductase gene (MTHFR), a reduction type coenzyme I quinone oxido-reductase gene (NQ01), an oncoprotein 53 gene (P53), an X-ray repair cross-complementing gene 1 (XRCC1) and a xeroderma pigmentosum complementary set A gene (XPA) as well as specific fluorescent probe pairs, fluorescent quantitative PCR regular components, PCR reaction components and the like. The kit of the invention estimates the individual genetic risk of esophageal cancer by synchronously testing the polymorphic locus genotypes of the genes closely related to the genetic risk of esophageal cancer.

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 esophageal cancer. P450 2E1 gene (CYP2E1), glutathione sulfur transferase M1 gene (GSTM1), 5,10-methylenetetrahydrofolate reductase gene (MTHFR), reduced coenzyme I quinone oxidoreductase gene (NQO1) , tumor protein 53 gene (P53), X-ray staggered complementation repair gene 1 (XRCC1), xeroderma pigmentosa complementation group A gene (XPA) genotypes of single nucleotide polymorphisms to evaluate individual esophageal cancer genetic predisposition. Background technique

[0002] Esophageal cancer, also called esophageal cancer, is a malignant tumor of the esophageal mucosal epithelium and esophageal glandular epithelium. The esophagus is the digestive tube connecting the pharynx and the stomach, and plays an important role in the digestive function of the human bo...

Examples

Embodiment 1

[0044] Example 1. Use of detection kits

[0045] Step 1: Extraction of DNA template

[0046] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0047] Step 2: Electrophoretic detection and typing

[0048] 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.

[0049] 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.

[0050] Then use electrophoresis technology to perform agarose gel electrophores...

Embodiment 2

[0058] Example 2. Application of detection kits for individual genetic susceptibility detection services for esophageal cancer

[0059] Step 1: DNA Extraction

[0060] 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.

[0061] Step 2: Genotyping Assays

[0062] Use the kit provided by the present invention to carry out PCR amplification and electrophoresis detection on the SNP site on the GSTM1 gene of the subject's genomic DNA, and perform PCR amplification and electrophoresis detection on the Ile462Val SNP site on the CYP1A1 gene, the Rsa I SNP site on the CYP2E1 gene, and the MTHFR gene The C677T SNP site on the NQO1 gene, the C609T SNP site on the NQO1 gene, the Arg72Pro SNP site on the P53 gene, the Arg194Trp SNP site on the XRCC1 gene, and the A23G SNP site on the XPA gene were detected by fluoresce...

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 esophageal cancer. P450 2E1 gene (CYP2E1), glutathione sulfur transferase M1 gene (GSTM1), 5,10-methylenetetrahydrofolate reductase gene (MTHFR), reduced coenzyme I quinone oxidoreductase gene (NQO1) , tumor protein 53 gene (P53), X-ray staggered complementation repair gene 1 (XRCC1), xeroderma pigmentosa complementation group A gene (XPA) genotypes of single nucleotide polymorphisms to evaluate individual esophageal cancer genetic predisposition. Background technique

[0002] Esophageal cancer, also called esophageal cancer, is a malignant tumor of the esophageal mucosal epithelium and esophageal glandular epithelium. The esophagus is the digestive tube connecting the pharynx and the stomach, and plays an important role in the digestive function of the human bo...

Examples

Embodiment 1

[0044] Example 1. Use of detection kits

[0045] Step 1: Extraction of DNA template

[0046] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0047] Step 2: Electrophoretic detection and typing

[0048] 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.

[0049] 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.

[0050] Then use electrophoresis technology to perform agarose gel electrophores...

Embodiment 2

[0058] Example 2. Application of detection kits for individual genetic susceptibility detection services for esophageal cancer

[0059] Step 1: DNA Extraction

[0060] 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.

[0061] Step 2: Genotyping Assays

[0062] Use the kit provided by the present invention to carry out PCR amplification and electrophoresis detection on the SNP site on the GSTM1 gene of the subject's genomic DNA, and perform PCR amplification and electrophoresis detection on the Ile462Val SNP site on the CYP1A1 gene, the Rsa I SNP site on the CYP2E1 gene, and the MTHFR gene The C677T SNP site on the NQO1 gene, the C609T SNP site on the NQO1 gene, the Arg72Pro SNP site on the P53 gene, the Arg194Trp SNP site on the XRCC1 gene, and the A23G SNP site on the XPA gene were detected by fluoresce...