A SNP marker and kit for screening high-risk groups of cardiac cancer
A cardia cancer and kit technology, applied in the fields of genetic engineering and oncology, can solve the problems of SNP sites and kits for high-risk groups of cardiac cancer, and achieve early screening convenience, ease of detection, high sensitivity and specific effect
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Embodiment 1
[0062] Example 1: Screening for detecting SNP sites related to susceptibility to cardia cancer
[0063] The inventors conducted genome-wide association analysis on peripheral blood DNA of 1528 patients with cardiac cancer and 6936 healthy people collected from areas with high incidence of cardiac cancer in my country (Linzhou City, Anyang City, Huixian City, Henan Province, etc.). There were 15 SNP loci that were significantly related to cancer. Among them, 1528 patients with cardiac cancer were pathologically diagnosed cardiac cancer patients, and 6936 healthy people were pathologically confirmed without cardiac cancer.
[0064] The specific operation of screening the 15 SNP loci significantly related to the incidence of cardia cancer is as follows:
[0065] 1. DNA extraction from peripheral blood: Extract DNA from the sample according to the instructions of the QIAGEN genomic DNA extraction kit. The specific steps are:
[0066] (1) Cell thawing: Take the blood sample out of ...
Embodiment 2
[0081] Example 2: Detection of susceptibility gene loci and screening of high-risk groups in asymptomatic people in 10,000 cases of cardiac cancer high-incidence areas
[0082] Select 10,000 asymptomatic people aged 50-60 in high-incidence areas of cardia cancer (Linzhou City, Anyang City, Huixian City, Changzhi City, etc.), and extract their peripheral blood DNA. The genotypes of the 15 SNP loci that were significantly associated with the above 10,000 asymptomatic patients were divided into high-risk and non-high-risk groups, and the two groups were followed up for 8 years and confirmed by gastroscopy and biopsy. The prevalence of cardia cancer in the two groups was analyzed, and statistical analysis was performed to determine the sensitivity and specificity of 15 SNP loci in screening high-risk populations for cardia cancer.
[0083] The specific operation steps are as follows:
[0084] 1. DNA extraction from peripheral blood: Extract the sample DNA according to the instruc...
Embodiment 3
[0131] Example 3: Production of a kit for screening high-risk and high-risk populations of cardia cancer
[0132] The production and operation process of the screening kit for high-risk and high-risk groups of cardia cancer is based on Affymetrix 6.0 chip detection and Sequenom MassARRAY genotyping technology. Specific amplification primers and specific extension primers for SNP sites that are significantly associated , dNTP mixture, MgCl 2 Solutions, PCR reaction buffers, enzyme digestion reaction buffers, deionized water, etc., these commonly used reagents are well known to those skilled in the art; in addition, there may also be standards and controls (such as standards and blanks for determining genotypes) control, etc.). The detection method of the kit includes: (1) DNA extraction from peripheral blood; (2) PCR amplification; (3) alkaline phosphatase treatment of PCR products; (4) single base extension; (5) resin purification; (6) ) chip spotting; (7) steps such as mas...
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