Method and kit for high-sensitivity detection of cancer related gene mutation

A detection method and high-sensitivity technology, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc.

Inactive Publication Date: 2017-09-15
GUANGDONG ASCENDAS GENOMICS TECH CO LTD
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  • Abstract
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Problems solved by technology

At present, blood EGFR mutation detection has not been widely carried out due to the above-mentioned limitations of next-generation sequencing, and the detection of low-frequency mutations in the EGFR gene requires a method with higher sensitivity and higher specificity

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  • Method and kit for high-sensitivity detection of cancer related gene mutation
  • Method and kit for high-sensitivity detection of cancer related gene mutation
  • Method and kit for high-sensitivity detection of cancer related gene mutation

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Embodiment Construction

[0094] The invention will be described in detail below. But it is not intended to limit the present invention.

[0095] The present invention detects cancer-related gene mutations by using labeled specific primers to first perform multiple PCR amplification on target genes, and then to perform high-throughput next-generation sequencing. Specifically, the following processing steps may be included.

[0096] 1. Design of labeled primers

[0097] Retrieve the DNA sequences of cancer-related genes (such as EGFR gene) mutation sites from online databases, compare them with software, design primers for these mutation regions, and add linker sequences and Unique Identifier (UID) sequences to the primer ends. After repeated tests, screening and verification, the labeled primers with high amplification efficiency and good specificity can be obtained correctly for low-frequency gene mutations in the samples to be detected.

[0098] Among them, a set of tagged primers is provided for ...

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Abstract

The invention provides a method and a kit for high-sensitivity detection of cancer related gene mutation. A tag primer group is designed for target gene, primers respectively undergo two-step amplification, and a next generation sequencing process is carried out. The method and the kit effectively eliminate false positive low-frequency mutation, and a detection result of the method and the kit has high sensitivity and high specificity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a high-throughput detection method and a kit for specifically and highly sensitively detecting low-frequency gene mutations, especially achieving good results in the detection of EGFR gene mutations. Background technique [0002] Plasma, as one of the important sources of liquid biopsy samples, contains more circulating DNA released from aging and dead cells. The patient’s plasma contains more free DNA, which is mainly produced by apoptotic and necrotic tumor cells. Its genetic characteristics are similar to those of The genomic DNA of tumors is similar, and the mutation forms include deletions, point mutations, and copy number gains. Therefore, plasma DNA has been used as a non-invasive detection sample and an important marker for early diagnosis of diseases, condition monitoring, curative effect and prognosis evaluation. [0003] Fine gene mutations can be obtained from the plasma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q2525/191C12Q2537/143C12Q2535/122
Inventor 赵鑫洪小龙甘海燕董志强吴涵杨呈勇
Owner GUANGDONG ASCENDAS GENOMICS TECH CO LTD
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