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Nucleic-acid mass spectrometry detection method for early screening of drive genes and susceptible genes of lung cancer

A susceptibility gene and driver gene technology, applied in the field of genetic testing, can solve the problems of incompletely including cutting-edge molecular targets, inability to achieve high-throughput detection and high-throughput analysis, and confidentiality of technical information.

Inactive Publication Date: 2017-12-26
武汉赛云博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The complexity analysis of molecular targets such as tumor development, drug resistance after targeted therapy, and metastasis mechanism, as well as the limitations of single-gene detection of multi-target variant genes in lung cancer patient samples, the cost of traditional genotyping detection methods High, unable to achieve high-throughput detection and high-throughput analysis, which determines the necessity and urgency of developing a multi-gene detection platform in clinical translational medical research
At present, the LungCarta kit can detect 214 mutation sites in 26 cancer-related genes, but the technical information is confidential, expensive, and does not fully include cutting-edge molecular targets, nor does it consider the differences in driver gene profiles in East Asia and Europe and the United States. Therefore, It is of great significance to establish a multi-gene detection method suitable for the Chinese population

Method used

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  • Nucleic-acid mass spectrometry detection method for early screening of drive genes and susceptible genes of lung cancer
  • Nucleic-acid mass spectrometry detection method for early screening of drive genes and susceptible genes of lung cancer
  • Nucleic-acid mass spectrometry detection method for early screening of drive genes and susceptible genes of lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0229] Example 1: Feasibility Analysis of Specific and Sensitive Gene Mutation Sites and SNPs Site Screening of Susceptibility Genes for Risk of Lung Cancer in Asian Population Individuals

[0230] The inventors of the present invention searched NCBI domestic and foreign genome-wide association study (genome-wide association study, GWAS) research literature and related patents (patent literature 6 and non-patent literature 7, etc., including but not limited to This), while searching the GWAS Catalog database and cancer-related databases such as COSMIC, CIViC, ICGC, and MYCANCER GENOME, screened out more than 1,000 positive samples from the Asian population or more than 500 positive samples from the Chinese population in the GWAS study. The inventors screened and evaluated the loci associated with the occurrence of lung cancer verified in the study, and selected 29 lung cancer hotspot mutations and 20 single nucleotide polymorphism loci that are significantly associated with the r...

Embodiment 2

[0248] Embodiment 2: Detection method of the present invention detects lung cancer cell lines

[0249] 1. Research object

[0250] Eight lung cancers with known mutation sites H460, PC9, H1650, H1975, A549, GLC82, HCC827, and H1299 (all of which can be purchased from ATCC) were selected for feasibility analysis of this detection method. ) (derived from the oral mucosa tissue extraction of normal people in Wuhan General Hospital of Guangzhou Military Region) as a negative control, water (H 2 0) as a blank control.

[0251] 2. Experimental steps

[0252] (1) Extract the DNA of the lung cancer cell line to obtain a DNA extract;

[0253] (2) Design the amplification primers of the gene mutation site and the SNPs site of the susceptibility gene used to assess the specificity and sensitivity of the individual suffering from lung cancer risk in Table 1 and Table 2 and the single base in Table 3 and Table 4 For extension primers, the amplification primers and single-base primers w...

Embodiment 3

[0289] Example 3: Detection of SNPs and mutation sites of a group of genes related to the early diagnosis of lung cancer from the peripheral blood of 26 pathologically diagnosed lung cancer patients

[0290] 1. Research object

[0291]In this study, peripheral venous blood samples from Wuhan General Hospital of Guangzhou Military Command were used to conduct genetic testing on the samples, and the number of cases was 26. The control group was a sample of healthy people from local health checkups.

[0292] 2. Experimental steps

[0293] The specific steps of applying the technology of the present invention to detect the SNPs and mutation sites of genes related to early diagnosis of lung cancer in peripheral blood samples are as follows:

[0294] (1) Obtain the subject's cfDNA sample using the Biotech Whole Blood Genomic DNA Rapid Extraction Kit

[0295] (a) Take 200 μl of collected peripheral blood and put it into a 1.5ml centrifuge tube. Add 200 μl of binding solution CB, ...

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Abstract

The invention belongs to the field of gene detection and particularly relates to a nucleic-acid mass spectrometry detection method for early screening of drive genes and susceptible genes of lung cancer. The nucleic-acid mass spectrometry detection method has the characteristics that by consideration of the difference of the lung-cancer gene spectra of the Asian populations and European-American populations, 29 lung-cancer hotspot mutations and 20 mononucleotide polymorphic sites significantly-related to suffering risk of the Asian populations are selected and combined, drive genes and susceptible genes of detection are advanced, multiple sites related to lung-cancer susceptibility and targeted-therapy drug resistance are introduced and are independent mutually without unbalanced linkage, therefore, site selection has representativeness, independence and risk-value accumulation, and can be used for evaluating the risk of individuals suffering from the lung cancer; and the detection technology is obvious in price advantage, higher in sensitivity at the aspect of lung-cancer early screening detection and larger in flux, can realize detection of multiple genes of single small samples to meet the maximum use of the small samples, and can realize translational research in clinical application.

Description

technical field [0001] The invention belongs to the field of gene detection, and in particular relates to a nucleic acid mass spectrometry detection method for early screening of lung cancer driver genes and susceptibility genes. Background technique [0002] Lung cancer is the number one cancer killer in China. Among the 653,000 new lung cancer patients in my country every year, non-small cell lung cancer (NSCLC) patients account for 85%. In contrast to the steady increase in survival rates for most cancers, progress in the treatment of lung cancer has been slow. Normally, the 5-year survival rate of lung cancer patients is 16%, and if diagnosed early, the 5-year survival rate of lung cancer patients rises to 52%. However, if metastases occur, this will drop to less than 4% by stage IV. Unfortunately, more than half of lung cancer patients are already stage IV lung cancer when they are diagnosed. Facing lung cancer is definitely a battle of life against time. Only by ear...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6886C12Q2600/106C12Q2600/156C12Q2531/113C12Q2521/525C12Q2533/101C12Q2565/627
Inventor 尚小云刘慧莹雷菁刁波
Owner 武汉赛云博生物科技有限公司
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