Kit for detecting epidermal growth factor receptor (EGFR) mutation by primer specific fluorescence polymerase chain reaction (PCR)

Abstract

The invention provides a kit for detecting the epidermal growth factor receptor (EGFR) mutation by a primer specific fluorescence polymerase chain reaction (PCR), in particular to the diagnosis of the EGFR gene mutation in tumor tissues and peripheral blood serums of tumor patients. Aiming at specific mutant sites relative to the curative effects of molecular targeted anti-cancer drugs, the kit is provided with a specific oligonucleotide primer sequence and a probe sequence, carries out fluorescence PCR detection respectively through a wild type PCR system and a mutation type PCR system on each sample to be detected and judges whether the samples have the mutation of EGFR19 exons and 21 exons or not through the magnitude of a difference value (delta Ct) of Ct values of two times of reaction. The kit can detect whether the specific sites of the EDFR have the mutation or not and can be used for the curative effect forecast of new molecular targeted anti-cancer drugs and the guidance of clinical individualized medicine application programs of the tumor patients.

Description

technical field [0001] The invention relates to a real-time fluorescent PCR detection kit, in particular to a kit for detecting common mutations of epidermal growth factor receptor (Epithelium Growth Factor Recptor, EGFR) by primer-specific real-time quantitative fluorescent PCR technology. Background technique [0002] Gefitinib (gefitinib, Iressa) EGFR tyrosine kinase inhibitor (EGFR-TKI), developed by AstraZeneca, is a new type of tumor-targeted therapy drug that effectively cuts off the epidermal growth factor receptor Send a signal into the cell, thereby hindering the development of tumors. Studies by scholars in the United States, Japan, and China in recent years have shown that somatic mutations of EGFR are associated with sensitivity to gefitinib in NSCLC patients. For tumors with EGFR mutations, the effective rate of Gefitinib is as high as 80%-90%, but for most tumors without EGFR mutations, Gefitinib has no effect. The EGFR protein tyrosine kinase functional reg...

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Problems solved by technology

However, on the one hand, the design of Taqman-MGB probes has strict requirements on the template sequence, and it is difficult to ensure that there are suitable probe sequences at all mutation positions; on the other hand, using the specificity of Taqman probes to detect mutations requires specific Design a pair of wild-type and mutant probes. For example, there are more than 20 kinds of deletion mutations of EGFR exon 19 reported in the literature. If 10 mutations are to be detected, 20 fluorescent probes are required, and Taqman-MGB probes Expensive, the use of multiple probes will greatly increase the detection cost

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