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Method for detecting genes of lung cancer and colorectal cancer based on NGS method

A technology for colorectal cancer and genetic detection, applied in the direction of microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of low cost and high sensitivity, and achieve the effect of improving the possibility and solving the problem of synthesis cost

Inactive Publication Date: 2018-05-01
上海仁东医学检验所有限公司
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Problems solved by technology

[0013] In addition, liquid biopsy techniques include digital PCR, ArmsPCR, etc., which have high sensitivity and low cost, and the disadvantage is that only a few known sites can be detected

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  • Method for detecting genes of lung cancer and colorectal cancer based on NGS method
  • Method for detecting genes of lung cancer and colorectal cancer based on NGS method
  • Method for detecting genes of lung cancer and colorectal cancer based on NGS method

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Embodiment Construction

[0025] The technical solution of the present invention is described in detail below. Those skilled in the art should understand that the technical solution of the invention can be modified or equivalently replaced, which should be covered by the scope of the claims of the present invention.

[0026] In this example, 15 lung cancer-related genes were selected for panel design, such as: EGFR, ALK, KRAS, BRAF, ROS1, RET, ERBB2, MET, AKT1, NRAS, PIK3CA, PTEN, DDR2, MAP2K1 (MEK1) and FGFR1, 12 genes related to colorectal cancer were also selected, such as: EGFR, KRAS, BRAF, ERBB2, MET, AKT1, NRAS, PIK3CA, PTEN, MAP2K1 (MEK1), FGFR1 and SMAD4.

[0027] The verification method adopted in the library building hybridization capture process of the present invention is as follows:

[0028] 1.1gDNA fragmentation and purification:

[0029] 1.1.1. Take 500ng according to the qubit concentration, add water to make up to 100μl, add covaris 130μl interrupted tube, set the program: 50W, 20%, ...

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Abstract

The invention provides a method for detecting genes of lung cancer and colorectal cancer based on an NGS method. The method comprises the following steps: carrying out sample collection, extracting DNA in a sample, carrying out molecular label library building, carrying out hybridizing with Panel, carrying out NGS sequencing, carrying out analysis aiming at data obtained after sequencing, and judging the gene type of the sample, wherein the Panel comprises SEQ ID No 1-SEQ ID No 618. The detection method provided by the invention belongs to a molecular diagnosis technology for realizing accurate treatment on patients suffering from lung cancer and colorectal cancer, liquid biopsy is supported, and the sensitivity achieves 0.1%.

Description

technical field [0001] The invention relates to a method for detecting cancer genes, in particular to a method for detecting genes of lung cancer and colorectal cancer based on an NGS method. Background technique [0002] High-throughput sequencing technology performs sequence determination on hundreds of thousands to several million DNA molecules at a time. It is possible to conduct a detailed and comprehensive analysis of the transcriptome and genome of a species, so it is also called Deep Sequencing (Deep Sequencing). The Illumina platform is now the mainstream next-generation sequencing platform, with a market share of over 90%. [0003] Compared with early gene detection technologies: Fluorescence in situ hybridization (FISH) has high sensitivity but is difficult to operate; traditional polymerase chain reaction (PCR) has advantages in ease of operation and rapidity, but technical principles limit It can only detect one or a few genes; although gene chip technology ca...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6869C12Q1/6886C12Q2600/156C12Q2535/122
Inventor 徐小红曹建军赵国栋
Owner 上海仁东医学检验所有限公司
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