Colorectal cancer cetuximab monoclonal antibody anti-drug trace amount DNA mutation detection method and device

A technology for cetuximab and colorectal cancer, which is applied in the directions of biochemical cleaning devices, biochemical equipment and methods, enzymology/microbiology devices, etc. It can increase the applicability, improve sensitivity, and flexible detection effect.

Active Publication Date: 2018-09-28
ZHEJIANG PROVINCIAL PEOPLES HOSPITAL +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These methods have the following limitations: (1) the detection results of surgical tissue detection in patients with recurrence and metastasis are inaccurate
Temporal heterogeneity caused by tumor evolution can lead to false negative results for some samples
At the same time, there is genomic heterogeneity at the spatial level within the tumor and among metastatic lesions in different par

Method used

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  • Colorectal cancer cetuximab monoclonal antibody anti-drug trace amount DNA mutation detection method and device
  • Colorectal cancer cetuximab monoclonal antibody anti-drug trace amount DNA mutation detection method and device
  • Colorectal cancer cetuximab monoclonal antibody anti-drug trace amount DNA mutation detection method and device

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Experimental program
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Embodiment

[0090] 1. Capture probe design:

[0091] According to the accumulated experience of the inventors in their work, the accumulated genetic variation information related to cetuximab resistance in colorectal cancer was integrated and analyzed, and the capture chip was designed to capture 11 hotspots related to cetuximab resistance. The entire exon region of the gene is designed into the capture chip, as shown in the table below.

[0092] Table 1: List of hotspot genes associated with cetuximab resistance

[0093] KRAS

NRAS

ERBB2

MET

EGFR

BRAF

PIK3CA

PTEN

MEK1

NF1

AKT1

[0094] 2. Cell-free DNA extraction method from peripheral blood

[0095] 2.1 Peripheral blood collection method

[0096] Streck blood collection tubes were used for blood collection, the blood collection volume was not less than 15ml, and the blood was mixed up and down immediately after blood collection, and the plasma separation was c...

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Abstract

The invention discloses a colorectal cancer cetuximab monoclonal antibody anti-drug trace amount DNA mutation detection method and a colorectal cancer cetuximab monoclonal antibody anti-drug trace amount DNA mutation detection device. The method comprises the following steps: extracting free DNA from a sample; constructing a library by using the free DNA; enriching the library; capturing the enriched library by using a capture probe to obtain captured DNA; sequencing the captured DNA to obtain the sequencing result; and comparing the sequencing result with a reference sequence and detecting amutation site. By the method, low-initial-quantity DNA enriched capture sequencing mutation detection is realized.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to cancer gene detection. More specifically, the invention relates to a method and device for colorectal cancer cetuximab drug-resistant trace DNA enrichment capture sequencing mutation detection. Background technique [0002] Colorectal cancer is the most common gastrointestinal tumor worldwide. According to the statistics of authoritative organizations, there are more than 1,400,000 new cases and nearly 700,000 deaths every year in the world, ranking the forefront of all malignant tumors in terms of morbidity and mortality [1]. In the United States, with the continuous popularization of early screening and the continuous improvement of perioperative treatment methods, the 5-year survival rate of localized and regional colorectal cancer has reached more than 70%, but the 5-year survival rate of metastatic colorectal cancer The survival rate is still about 13%-14%, suggesting that the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12M1/34C12M1/00C12Q1/6886
CPCC12Q1/6869C12Q1/6886C12Q2600/156C12Q2565/537
Inventor 杨柳叶松胡晓歌倪超常连鹏徐亚平易鑫杨玲
Owner ZHEJIANG PROVINCIAL PEOPLES HOSPITAL
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