Multiple enrichment method for detection of low-frequency mutations associated with targeted drugs in non-small cell lung cancer

Abstract

The present invention relates to a multiple enrichment detection method for non-small cell lung cancer targeted drug-related low-frequency mutations. The present invention uses 2235_2249del-15, 2236_2250del-15 and exon 20 with the highest frequency among deletion mutations on exon 19 of human EGFR gene The T790M mutation on exon 21 and the L858R mutation on exon 21 were used as targets, and primers, repressors, and TaqMan probes labeled with different fluorescence were designed for each mutation site, and the fluorescence during the amplification process was collected. Draw the amplification curve of the signal, compare the Cq value of the amplification curve with the threshold value determined by the standard curve, and judge whether there is a corresponding mutation in the sample. The system of the present invention is optimized for cfDNA, so that the system can achieve amplification and detection of target sequences under the condition of high template fragmentation degree and strong wild-type background. The method has the characteristics of sensitivity and multiplexing, which makes the detection operation more convenient, fast and accurate, and has great 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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