Digital PCR detection system for EGFR T790M site and application thereof

The invention provides a digital PCR detection system for an EGFR T790M site and application thereof. The detection system comprises a specific amplification primer of the EGFR T790M site and wild andmutant probes. By optimizing the concentration ratio of the primer probes in a PCR amplification system, the sensitivity and stability of the whole detection system are greatly improved. Meanwhile, by introducing a specific probe modification technology, the detection efficiency of the EGFR T790M site is improved, and particularly by using a nucleic acid locking technology to enhance combinationof the wild and mutant probes and nucleotides of corresponding sites, the detection efficiency of a low-concentration plasma free nucleic acid (cfDNA) sample is significantly improved. 本发明提供了一种EGFR T790M位点的数字PCR检测体系及其应用。所述检测体系包含EGFR T790M位点的特异性扩增引物和野生型、突变型探针，通过优化PCR扩增体系中引物探针的浓度比例，使得整个检测体系的灵敏度和稳定性大幅度提高。同时通过引入特异性的探针修饰技术增强EGFR T790M位点的检测效率，尤其是使用锁核酸技术增强野生型和突变型探针与对应位点核苷酸的结合，显著的提高了低浓度血浆游离核酸(cfDNA)样本的检测效率。