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Colorectal cancer susceptibility diagnostic kit and application of SNP (single nucleotide polymorphism) in preparation of diagnostic kit

A technology of colorectal cancer, kit, applied in the field of biomedicine

Inactive Publication Date: 2014-12-24
PEOPLES HOSPITAL PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The pathogenesis of colorectal cancer has not been fully elucidated, but etiological studies have shown that the occurrence of most colorectal cancers is a multi-step, multi-stage complex process with the combined effects of environmental factors and genetic factors

Method used

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  • Colorectal cancer susceptibility diagnostic kit and application of SNP (single nucleotide polymorphism) in preparation of diagnostic kit
  • Colorectal cancer susceptibility diagnostic kit and application of SNP (single nucleotide polymorphism) in preparation of diagnostic kit
  • Colorectal cancer susceptibility diagnostic kit and application of SNP (single nucleotide polymorphism) in preparation of diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Identification of Genomic SNPs Sites Related to Human Colorectal Cancer Susceptibility

[0037] 1. Research object

[0038] This study is divided into three parts: GWAS preliminary screening stage, first stage verification and second stage verification. In the initial screening stage of GWAS, samples from Beijing area (mainly from Peking University People's Hospital and Peking University Cancer Hospital) were used, with 932 cases and 1006 cases in the control group. The first phase of validation used samples from central China (Jiangsu and Shanghai), including 1759 cases and 1875 controls. The second phase of validation included 943 cases and 1838 controls from Beijing, Shandong, and Northeast China. All cases were hospital-based sources of Han population, colorectal cancer confirmed by histopathology or cytology. The control group came from local health checkups and healthy population samples from the China Bone Marrow Bank, and was matched with cases acco...

Embodiment 2

[0089] Example 2 Sequenom MassArray detects the genotype of rs12522693 or rs17836917

[0090] (1) The genomic method for extracting samples is the same as in Example 1.

[0091] (2) Amplify the DNA region containing the target site. The primer sequence used is as follows: For the rs12522693 site, the amplification primer sequence is: forward primer 5'-ACGTTGGATGCAAGATGACTCTAACTACC-3', reverse primer 5' -ACGTTGGATGCCTATAAGTGCAAAGGAGAG-3'; for the rs17836917 site, the amplification primer sequence is: forward primer 5'-ACGTTGGATGCCCAGCCTGGTTTGTATTGT-3', reverse primer 5'-ACGTTGGATGTCCTGCCTTCTTAATTCTGC-3'.

[0092] (3) Use shrimp alkaline phosphatase SAP to remove free dNTPs in the system, and purify the PCR product;

[0093] (4) Single-base extension reaction: The PCR product after SAP treatment undergoes base-specific extension reaction, and the molecular weight of the extension products of different alleles is different, that is, the difference in the molecular weight of the ...

Embodiment 3

[0097] Example 3 Detection of the genotype at the rs12522693 or rs17836917 site by direct sequencing

[0098] (1) The genomic method for extracting samples is the same as in Example 1.

[0099] (2) PCR amplification a. Perform PCR amplification on the sample DNA (for example, Takara PCR Thermal Cycler PCR amplification instrument from TaKaRa Company, Japan can be used). The primer sequence used is as follows: for the rs12522693 site, the amplification primer sequence is as follows: forward primer 5'-ACGTTGGATGCAAGATGACTCTAACTACC-3', reverse primer 5'-ACGTTGGATGCCTATAAGTGCAAAGGAGAG-3'; for rs17836917 site Say, the amplification primer sequences are as follows: forward primer 5'-ACGTTGGATGCCCCAGCCTGGTTTGTATTGT-3', reverse primer 5'-ACGTTGGATGTCCTGCCTTCTTAATTCTGC-3'. Use 50μl PCR reaction system for gene amplification of rs12522693 or rs17836917, containing 1×PCR buffer, 1.5mM MgCl 2 , 100-150ng of genomic DNA, both upstream and downstream primers are 0.5μM, dNTP is 0.2mM, and ...

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Abstract

The invention discloses a colorectal cancer susceptibility diagnostic kit and an application of SNP (single nucleotide polymorphism) in preparation of the diagnostic kit. The SNP sites are rs12522693 and rs17836917. By means of genome-wide association study, the inventors prove remarkable correlation between an allele at the rs12522693 site G and an allele at the rs17836917 site A and colorectal cancer susceptibility through a screening stage, first stage verification and second stage verification. By means of detecting the genotypes of the rs12522693 site and / or the rs17836917 site, the colorectal cancer susceptibility of a testee can be predicted, and thus early prevention and treatment of the disease can be promoted. The colorectal cancer susceptibility diagnostic kit can be used for detecting the genotypes of the rs12522693 site and / or the rs17836917 site by using known technologies in the field so as to judge whether the testee has colorectal cancer susceptibility or not.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a colorectal cancer susceptibility diagnostic kit and the use of SNP in preparing the colorectal cancer susceptibility diagnostic kit. Background technique [0002] Colorectal cancer is the third most common malignant tumor in both men and women. According to the 2013 American Cancer Statistics Report, there were 103,170 new cases of colon cancer in the United States in 2012, and 40,290 cases of rectal cancer, with a mortality rate of 50,830. In my country, with the changes in lifestyle and eating habits in recent years, the incidence and death of colorectal cancer have risen rapidly. According to the statistical results in 2011, colorectal cancer ranks third in the incidence of malignant tumors in my country and fourth. The morbidity and mortality rate of malignant tumors are close to the world level at the same time, and it has become a public health problem that seriously threatens the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/156
Inventor 王杉姜可伟程京胡志斌孙义民沈洪兵叶颖江吕亮张继准
Owner PEOPLES HOSPITAL PEKING UNIV
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