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Method for detecting tumor-related polygene mutation by utilizing high-throughput sequencing

A tumor-related, high-throughput technology, used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., to achieve stable and reliable quality, short time consumption, and good integrity.

Pending Publication Date: 2021-03-02
WUHAN ADICON CLINICAL LAB
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional gene mutation detection methods, such as Sanger sequencing, pyrosequencing, and real-time fluorescent PCR, can only detect a single gene, or some exon mutations of a single gene, and have limitations in the detection of complex mutation types

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  • Method for detecting tumor-related polygene mutation by utilizing high-throughput sequencing
  • Method for detecting tumor-related polygene mutation by utilizing high-throughput sequencing
  • Method for detecting tumor-related polygene mutation by utilizing high-throughput sequencing

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Experimental program
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Embodiment Construction

[0024] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0025] 1.1 Sample processing

[0026] All paraffin samples must first be sectioned by pathology, stained with HE, and examined by pathology microscope. After pathological microscopic examination, it is required to contain tumor tissue and the sample volume should be >100 cells before entering the subsequent sample preparation and nucleic acid extraction process. After the tissue enters the laboratory, press figure 1 Process processing.

[0027] 1.1.1 DNA extraction (column extraction method)

[0028] 1.1.1.1 Take 5-8 paraffin sections (5-10μm thick, 1×1cm2 size), remove excess paraffin with a clean blade, cut the sample (<20mg) into as small pieces as possible, and transfer to a 1.5ml centrifuge tube middle. Removing excess paraffin or cutting the sample into as small pieces as possible with scissors or blades is beneficial for s...

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Abstract

The invention discloses a method for detecting tumor-related polygene mutation by using high-throughput sequencing. The method comprises the following steps of (1) extracting DNA by using a column extraction method; (2) constructing a database, namely breaking DNA, repairing a tail end and adding an A tail; then connecting a joint, carrying out first-step magnetic bead purification, then carryingout UDI joint primer PCR amplification, and then carrying out second-step magnetic bead purification; (3) hybrid capture; the method comprises steps of database hybridization, database capture and cleaning, PCR amplification and magnetic bead purification. (4) sequencing; and (5) biological information analysis and result interpretation. The method is advantaged in that the sample mutation information, MSI and other information are obtained by sequencing captured fragments and analyzing subsequent biological information, the relationship between multiple drugs and gene variation is analyzed, gene detection is helped for tumor patients, and the kit is especially suitable for middle and advanced stage patients with recurrence and metastasis.

Description

technical field [0001] The invention relates to a method for detecting tumor-related multi-gene mutations by using high-throughput sequencing. Background technique [0002] Since the 20th century, cancer has become one of the major threats to human health. According to the 2016 data from the National Cancer Center, in 2015, the number of new cancer cases in China is expected to be about 4.29 million, and the total number of new deaths will be about 2.81 million. Lung cancer is the most common among many cancers and is the leading cause of death in cancer patients. Cancers of the stomach, esophagus and liver also rank among the top causes of cancer death." [0003] Cancer initiation, progression, metastasis, treatment response and drug resistance may result from the continuous accumulation of mutations in tumor cell clones. The mutations in tumor cells range from thousands to hundreds of thousands. However, most mutations have no substantial role in cancer progression and ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6869
CPCC12Q1/6886C12Q1/6869C12Q2600/156C12Q2531/113C12Q2523/308C12Q2535/122
Inventor 周桂兰
Owner WUHAN ADICON CLINICAL LAB
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