Multiple enrichment detection method of low-frequency mutation relevant to non-small cell lung cancer target medicine

Abstract

The invention relates to a multiple enrichment detection method of low-frequency mutation relevant to a non-small cell lung cancer target medicine. Four mutations of 2235_2249del-15 and 2236_2250del-15 which are highest in frequency in deletion mutations on an exon 19 of a human EGFR gene, a T790M mutation on an exon 20, and an L858R mutation on an exon 21 are used as targets, primers, blockers and TaqMan probes marking different fluorescence are respectively designed in accordance with each mutation site, an amplification curve is drawn through collecting fluorescence signals in the amplification course, the Cq value of the amplification curve and a threshold determined through a standard curve are compared, and whether samples have corresponding mutations or not is judged. The system disclosed by the invention is optimized in accordance with cfDNA, so that the system can realize amplification and detection on a target sequence under the situation that the formwork gragmentation degree is high and the wild type background is powerful. The method concurrently has the characteristics of being sensitive and multiple, the detection operations are convenient, quick and accurate, and the method has large application value.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a multiple enrichment detection method for non-small cell lung cancer targeting drug-related low-frequency mutations based on qPCR technology. Background technique [0002] At present, the morbidity and mortality of lung cancer rank first among all kinds of cancers in the world. According to World Health Organization estimates, in 2012, there were 1.8 million new cases worldwide, accounting for 12.9% of all cancers; 1.59 million deaths, accounting for 19.4% of all cancers. Among them, lung cancer is the cancer with the highest incidence rate in men. China is a country with a high incidence of lung cancer (Ferlay J, Soerjomataram I, Ervik M, et al. GLOBOCAN 2012v1.0, cancer incidence and mortality worldwide: IARC cancer base no.11 [Internet]. International Agency for Research on Cancer, Lyon [J]) . According to the 2015 World Health Organization classif...

AI Technical Summary

Problems solved by technology

dPCR technology can detect and quantify low-frequency mutations more accurately, but it relies on complex instruments, low throughput, high detection costs, and complicated data processing, so it is mainly used in scientific research

The next-generation sequencing technology also requires complex instruments, and the inherent error rate of the technology seriously affects the specificity of detection. In addition, the higher cost and longer detection time also hinder its application.

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