Primers, probes and kit for detecting 19-Del mutation of epidermal growth factor receptor (EGFR) gene

Abstract

The invention discloses nucleic acid sequences and a kit for detecting 19-Del mutation of an epidermal growth factor receptor (EGFR) gene and belongs to the technical field of molecular diagnosis. Theprimers, the probes and the kit for detecting 19-Del mutation of the EGFR gene can detect 53 common mutation types (covering 99% of mutation occurrence rate or above) of EGFR-TKIs targeting drugs forclinical NSCLC patients with high sensitivity and high specificity. The invention also discloses the kit for detecting 19-Del mutation of the EGFR gene. The kit can realize quantitative detection ofmutation of 19-Del of the EGFR gene in one hole conveniently, quickly, sensitively, specifically and accurately, can be used as the reference basis of EGFR-TKIs targeting drugs and can be used for dynamically monitoring curative effects of drugs in real time.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and in particular relates to a primer, a probe and a kit for detecting EGFR gene 19-Del mutation. Background technique [0002] In recent years, targeted drugs such as epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) have become important in advanced non-small cell lung cancer (NSCLC). one of the treatment modalities. NCCN, ESMO, CSCO and other domestic and foreign tumor diagnosis and treatment and clinical practice guidelines recommend that all NSCLC patients with adenocarcinoma components must undergo EGFR gene mutation detection to guide the selection of EGFR-TKI targeted therapy. [0003] A large number of clinical studies have found that patients with non-small cell lung cancer cells carrying EGFR gene sensitive mutations, use EGFR tyrosine kinase inhibitors (EGFR-TKI) such as Iressa (gefitinib), Tarceva (erlotinib) Niger), compared with standard first-li...

AI Technical Summary

Problems solved by technology

Some researchers in the early stage have developed a variety of 19-Del multiplex detection systems and methods based on ARMS, NGS and digital PCR technologies, but the systems and methods they developed have their limitations, and the methods based on NGS technology are relatively comprehensive in detecting mutations. , but the sensitivity is insufficient, the operation is complicated, difficult, and costly, which cannot meet clinical requirements; the sensitivity of ARMS technology is also low, and there are only a few to more than 20 detection sites, and there are many mutations that cannot be detected, resulting in missed detection It will bring serious consequences to the patient’s medication. The amplification block method is relatively harsh on the sample volume of clinical samples, and must be diluted to a certain concentration range. Too much sample volume will easily cause false positives, and too low will reduce its sensitivity and cause leakage. detection; ddPCR technology has high sensitivity and accurate quantification, but only 10-30 kinds of mutations can be detected in the current relevant literature and patents, covering 60-95% of the mutation incidence rate, and more than 1 / 3 of the types of mutations, about 5-15% of 19-Del mutation patients cannot be detected, and some missed detection cannot be avoided

Therefore, none of the existing products can effectively meet the current needs of clinical NSCLC patients for precise personalized medicine and companion diagnostics

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