Method and kit for detecting non-small cell lung cancer drive gene mutation spectrum, and application

Abstract

The invention discloses a method and a kit for detecting a non-small cell lung cancer drive gene mutation spectrum, and an application. The method comprises the following steps of: designing 15 pairs of amplification primers for amplifying the exon segments of the seven related genes of non-small cell lung cancer, dividing the amplification primers into 6 groups, and preparing an amplification primer mixed solution, performing multiple PCR (polymerase chain reaction) amplification on the to-be-detected samples by the amplification primer mixed solution respectively, and then performing enzymatic digestion; and designing 39 extension primers used for detecting hotspot mutation sites, dividing the extension primers into 6 groups corresponding to the amplification primer mixed solution, and preparing an extension primer mixed solution, performing extension reaction on the digested PCR product, then performing enzymatic digestion, performing capillary electrophoresis on the obtained product, and making a result judgment via software analysis. The kit provided by the invention comprises the amplification primers for amplifying the exon segments of the seven related genes of non-small cell lung cancer, and the extension primers used for detecting hotspot mutation sites. The method and the kit provided by the invention are simple, high in flux, and short in time consumption.

Description

technical field [0001] The invention relates to a method for detecting the mutation spectrum of driving genes of non-small cell lung cancer, in particular to a method, kit and application for detecting the mutation spectrum of driving genes of non-small cell lung cancer. Background technique [0002] The morbidity and mortality of lung cancer rank first among tumors. The traditional treatment method is to select the chemotherapy regimen based on the patient's clinical characteristics and pathological type, but the progression-free period (PFS) is only 6 months. Targeted therapy for lung cancer has made remarkable progress in recent years, the most prominent example being: small molecule tyrosine kinase inhibitors targeting epidermal growth factor (EGFR) in patients with non-small cell lung cancer with mutant EGFR It showed significant efficacy, and the PFS reached 9-13 months. Subsequently, crizotinib targeting the EML4-ALK fusion gene also showed the same clinical efficac...

AI Technical Summary

Problems solved by technology

[0008] At present, the common detection methods of gene mutations include: direct sequencing method, ARMS method, HRM method, DHPLC method, etc. The common disadvantage of these methods is that only one mutation or a mutation of a gene can be detected at a time, and the detection throughput is low, so they can only be detected one by one. Screening for mutation status of driver genes, patients wait longer for results

At the same time, due to the difficulty in obtaining materials for advanced patients and the small sample size, it often cannot meet the needs of the above methods for detecting gene mutation spectrum.

Although the deep sequencing method can meet the clinical needs in terms of throughput, it cannot be popularized in clinical applications due to its high price.

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