Detection method of EGFR T790M and C797S cis mutation

Abstract

The invention discloses a detection method of EGFR cis mutation. Based on a Blocker PCR technology, different to-be-detected mutation sites are separately selected on a sense strand and antisense strand of a wild-type EGFR gene to design a corresponding Blocker, and the Blocker can be separately combined with the to-be-detected mutation sites of the sense strand and antisense strand of the wild-type EGFR gene to form a stable hybrid to prevent an upstream primer and a downstream primer from amplifying the wild-type EGFR gene which is subjected to monotonous mutation or trans-mutation. The method is simple and convenient, EGFR cis-configuration mutation can be effectively distinguished from trans-configuration mutation, the sensitivity is high, even 0.1% mutation in a peripheral blood sample can still be detected, and the detection method has a very high application value in clinical diagnosis of drug-resistant mutation of TKI.

Description

technical field [0001] The invention belongs to the field of gene detection, and in particular relates to a method for detecting EGFR cis-mutation based on Blocker PCR technology. Background technique [0002] EGFR is the expression product of the proto-oncogene c-erbB1 and a member of the epidermal growth factor receptor (HER) family. EGFR is a transmembrane tyrosine kinase receptor, and the activation of the receptor kinase domain is related to various signal transduction pathways such as cancer cell proliferation, metastasis and apoptosis. EGFR mutations mainly occur in the first four exons (18-21) of the intracellular tyrosine kinase (TK) region, and more than 30 mutations in the TK region have been discovered so far. The T790M substitution mutation on exon 20 is a drug-resistant mutation of a first-generation EGFR tyrosine kinase inhibitor (Tyrosine Kinase Inhibitor, TKI). TKI has always been an important drug in the targeted therapy of EGFR-mutant NSCLC, but most pat...

AI Technical Summary

Problems solved by technology

The third-generation EGFR-TKI Tagrisso/Tagrisso (also known as Osimertinib/Osimertinib, AZD9291) is an irreversible selective TKI, a new generation of targeted drugs that can effectively deal with the T790M mutation, and is aimed at overcoming the secondary drug resistance of T790M Here comes hope, but eventually drug resistance

[0004] Under the existing technology, next-generation gene sequencing technology (NGS) can distinguish the mutation configuration of C797S and T790M, while other PCR techn

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