Composition, kit and method for detecting EGFR mutation

Abstract

The invention discloses a composition, a kit and a method for detecting EGFR mutation. The composition includes forward primers, reverse primers and wild-type EGFR gene binding molecules, and the combination of the forward primers and the reverse primers can obtain amplified products containing EGFR mutation sites. The forward primers include 5' end regions, 3' end regions and intermediate regions. The two-side sequences of the 5' end regions can complement each other to form a stem-loop structure. The first ends of the 5' end regions have fluorescent groups and the second ends have quenchinggroups. The last bases of the 3' end regions complement mutant bases to be detected. The reverse primers include tag sequences located at the 5' end and binding regions that completely complement downstream sequences at the mutation sites to be detected. The wild-type EGFR gene binding molecules is set to be able to be specifically recognized and is combined to the mutation sites to be detected ofwild-type EGFR genes. The sensitivity and specificity of EGFR detection are improved.

Description

technical field [0001] The invention relates to gene detection, in particular to a composition, a kit and a method for detecting EGFR mutation. Background technique [0002] With the rapid development of translational medicine, the treatment of patients with advanced non-small cell lung cancer (NSCLC) has shifted from the era of chemotherapy to the era of molecular targeted therapy, in which epidermal growth factor receptor (EGFR) is the most important driver of lung cancer. EGFR-TKIs are small molecule inhibitors targeting this target, and advanced NSCLC patients with activating mutations in the EGFR gene can benefit significantly from EGFR-TKIs. However, almost all patients taking EGFR-TKI will eventually develop drug resistance. Early studies have found that about 50% of drug-resistant patients have a missense mutation in codon 790 of exon 20 of EGFR (ie T790M mutation), and it is related to EGFR -TKI curative effect is closely related, how to detect T790M gene mutation ...

AI Technical Summary

Problems solved by technology

For example, although the common ARMS method has a sensitivity of about 1%, it cannot detect samples with lower mutation frequencies.

The ddPCR method can quantitatively detect the T790M mutation, and the sensitivity can reach 0.4% or even higher, but the consumables are expensive and the operation is complicated

NGS is a Sequencing by Synthesis (SBS) technology with high throughput, high depth, and high price. It is suitable for multi-gene and multi-locus detection, but not suitable for detection of a small number of loci.

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