Method for monitoring and controlling drug resistance of colorectal cancer patient to panitumumab/cetuximab through ddPCR technology

A technology of cetuximab and panitumumab, applied in the medical and biological fields, can solve the problems of difficult to detect the type and sequence of cell-free DNA, interference of blood-derived DNA, and low content of cell-free DNA in plasma

Pending Publication Date: 2016-08-10
SHANGHAI BIOTECAN PHARMA +2
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Problems solved by technology

[0008] However, due to the extremely small amount of plasma cell-free DNA in the blood and the interference of a large amount of blood-derived DNA, it is still difficult to accurately isolate and purify DNA with high-efficiency DNA enrichment methods and high-sensitivity detection methods (such as sequencing, digital PCR). and reliably detect the specific types and sequences of tumor-associated cell-free DNA
For example, it has been reported in the literature that although the average content of circulating cell-free D

Method used

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  • Method for monitoring and controlling drug resistance of colorectal cancer patient to panitumumab/cetuximab through ddPCR technology
  • Method for monitoring and controlling drug resistance of colorectal cancer patient to panitumumab/cetuximab through ddPCR technology
  • Method for monitoring and controlling drug resistance of colorectal cancer patient to panitumumab/cetuximab through ddPCR technology

Examples

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Embodiment 1

[0190] Example 1 A method of detecting KRAS gene mutation in serum / plasma by ddPCR technology to monitor colorectal cancer patients' resistance to panitumumab / cetuximab

[0191] (1) Before starting panitumumab / cetuximab therapy in a patient with colorectal cancer, 4 mL of peripheral blood was collected, plasma / serum was separated, and free DNA in the plasma / serum was extracted. High-concentration / high-purity DNA was obtained through sample dissolution, enrichment, column elution, and finally AVE elution, and finally the fragment size (<313bp) was determined by agarose gel electrophoresis to ensure the extraction quality.

[0192] (2) Reaction system preparation: a. Probe and primer design: The primers used in the experiment were specific primers containing KRAS codon 12 / 13 sites, and the probes used included a mutant-specific probe and wild-type probe for each site. specific probes. The mutant probes are labeled with FAM fluorescence, and the wild-type probes are labeled with...

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Abstract

The invention provides a method for monitoring and controlling drug resistance of a colorectal cancer patient to panitumumab/cetuximab through ddPCR technology, and an application of the method to monitoring and controlling of panitumumab/cetuximab in a process of treating colorectal cancer. The method comprises: firstly extracting DNA from peripheral plasma of a colorectal cancer patient, then detecting a mutation situation of a drug-resistant mutation site G13D of KRAS in the DNA through ddPCR technology, and finally judging if the patient produces drug resistance according to the mutation situation.

Description

technical field [0001] The invention relates to the fields of medicine and biotechnology, in particular to a molecular detection technology that can be used to guide tumor treatment, especially a method for highly sensitive detection of the G13D gene mutation at the KRAS gene drug resistance mutation site in peripheral plasma and its role in monitoring colorectal cancer Patients' resistance to panitumumab / cetuximab and other aspects. Background technique [0002] With the development of biomedical technology, molecular targeted drugs have been widely used in the treatment of tumors. Compared with conventional chemotherapy, targeted therapy has the advantages of better effect and less side effects. Clinical application results show that some patients will develop drug resistance to targeted drugs. Research by the American Cancer Society shows that more than 90% of tumor patients are affected by drug resistance to varying degrees during treatment. The drug resistance of tumo...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 丁飞飞许骋沈波易静张桢珍吴云鸣楼敬伟张有成王青兵刘涛刘冰
Owner SHANGHAI BIOTECAN PHARMA
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