Kit for detecting gastric cancer risk gene and using method thereof

A gene detection and kit technology, which is applied in the field of gastric cancer risk gene detection kits, can solve the problems of increasing the risk of gastric cancer and achieve the effects of convenient and accurate application, pollution prevention and simple operation

Pending Publication Date: 2019-06-25
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In patients with HP infection in Hexi area, IL-10-819C allele IL-10-592C allele can increase the risk of gastric cancer

Method used

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  • Kit for detecting gastric cancer risk gene and using method thereof
  • Kit for detecting gastric cancer risk gene and using method thereof
  • Kit for detecting gastric cancer risk gene and using method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Such as figure 1 As shown, a gastric cancer risk gene detection kit of the present invention includes detection reagents for detecting one or more of 11 single nucleotide polymorphism sites in 9 genes, 11 of 9 genes The single nucleotide polymorphism sites are specifically: T→C of COX-2 gene rs5273 and G→C variation of rs20417, A→G variation of IL-10 gene rs1800896; C→G variation of P53 gene rs1042522; T→C of PARP-1 gene rs1136410 and A→G variation of rs3219145; A→G variation of PLCE1 gene rs2274223; T→C variation of RKAA1 gene rs13361707; C→T variation of PSCA gene rs2294008; G→G variation of SOD1 gene rs4998557 A variation; T→C variation of SOD2 gene rs4880.

[0041] The detection reagents include: PCR reagents and genotype determination reagents.

[0042] Wherein, the genotype determination reagents are Taqman probe fluorescent PCR reagents, Snapshot reagents, reagents for fluorescent PCR method, reagents for constant temperature amplification-fluorescence method, ...

Embodiment 2

[0108] The difference between this example and Example 1 lies in that the methods for analyzing and determining the genotype of the target sequence are different.

[0109] 3. Analysis and determination of target sequence genotype

[0110] Generation Sequencing

[0111] Nucleotide sequence determination:

[0112] 1. Glue recovery:

[0113] 1.1 Add Binding Buffer 30ul to the tube containing gel recovery and incubate at 55°C for 7min until it completely dissolves;

[0114] 1.2 After the gel is melted, take out the column tube, mark it, transfer the liquid into the column tube, centrifuge at 10000rpm / 1min, pour off the supernatant after completion, add 700ulSpw, let stand for 2-3min, centrifuge at 10000rpm / 1min, pour Drop the supernatant;

[0115] 1.3 Repeat the above 1.2 operation once;

[0116] 1.4 After pouring off the supernatant, the column tube is idling at 1000rpm / 1min;

[0117] 1.5 Add 15ulElution Buffer to the center of the column, let it stand for 2-3min, and centr...

Embodiment 3

[0130] The difference between this example and Example 1 lies in that the methods for analyzing and determining the genotype of the target sequence are different.

[0131] 3. Analysis and determination of target sequence genotype

[0132] Taqman probe fluorescent PCR method

[0133] reaction system:

[0134] Component

Volume

Final Concentration

dna

1pg-1ug

Forward primer (10uM)

0.4ul

0.2uM

Reverse primer (10uM)

0.4ul

0.2uM

Probe(10uM)

0.4ul

0.2uM

qPCR SuperMix

10ul

1×

Passive reference Dye (50x)

0.4ul

1×

Nuclease-free Water

Variable

-

Total volume

20ul

-

[0135] Reaction conditions:

[0136]

[0137] Amplification primers and probes:

[0138]

[0139]

[0140] In the present invention, other methods can also be used for the analysis and determination of the target sequence genotype. Such as:

[0141] 1. Chip detection based...

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PUM

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Abstract

The invention discloses a kit for detecting gastric cancer risk gene, which comprises a detection reagent for detecting one or more of 11 mononucleotide polymorphism sites in 9 genes, concretely for detecting COX-2 gene rs5273 and rs20417 mutation; IL-10 gene rs1800896 mutation; P53 gene rs1042522 mutation; PARP-1 gene rs1136410 and rs3219145 mutation; PLCE1 gene rs2274223 mutation; RKAA1 gene rs13361707 mutation; PSCA gene rs2294008 mutation; SOD1 gene Rs4998557 mutation; and SOD2 gene rs4880 mutation. The kit can be used for detecting a plurality of mutant genes in a sample of an individualto-be-tested, the occurrence of gastric cancer can be monitored or diagnosed at an early stage, and a reference for clinical diagnosis and treatment can be provided.

Description

technical field [0001] The present invention relates to the field of disease risk gene detection, in particular to a gastric cancer risk gene detection kit and its application method. Background technique [0002] Gastric cancer is a common digestive system tumor in my country, accounting for the first place in the death of malignant tumors. It is currently known that the occurrence of gastric cancer is related to basic gastric diseases (such as gastritis, gastrectomy, etc.), Helicobacter pylori (HP) infection, living habits such as high-salt diet, and genetic susceptibility factors. Studies have shown that 35%-60% of gastric cancer is related to HP infection, but only some of the HP infected people suffer from gastric cancer. Genetic factors may be another important factor in the occurrence of gastric cancer. The phenomenon of family clustering of gastric cancer and the fact that only a small number of people are affected by the same exposure environment indicate that the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
Inventor 李玉民刘涛
Owner LANZHOU UNIVERSITY
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