DdPCR detection method for lung cancers EGFR L858R and 19Del and application of ddPCR detection method

Abstract

The invention relates to the technical field of medicines and biology and particularly relates to a ddPCR detection method for lung cancers EGFR L858R and 19Del and application of the ddPCR detectionmethod. The detection method comprises the steps of separating hemocyte and plasma of peripheral blood, carrying out cfDNA extraction by virtue of a two-step purification method, respectively designing two probes and one pair of primers aiming at lung cancer-relevant EGFR L858R loci and EGFR 19# exon deletion loci, carrying out ddPCR detection by taking extracted cfDNA as a template, and finally carrying out data analysis on an obtained amplification product, so as to obtain the proportion of EGFR sensitively-mutated L858R and 19Del. The application comprises the application of the ddPCR detection method in the treatment of tumor patients, medication-relevant genetic locus mutation information is provided for the tumor patients in the treatment process of the terminal lung cancer, and important reference frame is provided for the individualized accurate medication guidance, pesticide effect evolution and treatment effect monitoring of the lung cancers.

Description

technical field [0001] The invention relates to the fields of medicine and biotechnology, in particular to a ddPCR detection method and application of lung cancer EGFR L858R and 19Del. Background technique [0002] Lung cancer is the most common primary malignant tumor of the lung. The vast majority of lung cancers originate from the bronchial mucosal epithelium, so it is also called bronchial lung cancer. Over the past 50 years, the morbidity and mortality of lung cancer have risen rapidly in countries all over the world, especially industrially developed countries. It is the malignant tumor with the highest morbidity and mortality in the world. The 5-year survival rate is only 16.8%. The number of patients has reached 1.6 million. Lung cancer is also the leading cause of death of cancer patients, and 1.4 million people die of lung cancer every year. [0003] At present, the traditional lung cancer treatment methods include: surgery, radiotherapy and chemotherapy. Althoug...

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Problems solved by technology

[0009] The existing detection methods for lung cancer have the following defects and deficiencies: First, the traditional detection optimization method mainly relies on the experience of technicians to interpret the results, which cannot directly, objectively and accurately reflect the detection results, nor can it perform absolute quantification of DNA; , the existing detection optimization method based on lung cancer EGFR L858R and 19Del usually has high requirements on the total amount and quality of cfDNA in the sample, and is easily affected by a large amount of blood-related DNA and response inhibitors; thirdly, the existing detection method based on lung cancer EGFR L858R and 19Del The detection optimization method of the method is cumbersome, and the detection sensitivity is low, and there will be a certain amount of false positive detection results, especially the cfDNA false positive detection rate in plasma samples is higher, because cfDNA is mainly derived from benign cells of germline origin. The proportion of ctDNA is relatively low; fourth, there are literatures showing that the addition of carrierRNA in the cfDNA extraction process does not increase the total amount of cfDNA extraction, and it is likely to cause large fragments of gDNA to aggregate

Fourth, the steps of liquid biopsy are cumbersome, the cycle is long, and the cost is high, so it is not suitable for the detection of single or two sites

[0010] In summary, circulating tumor DNA-based detection technology has become the focus, but there is still a lack of low-input, high-sensitivity, and high-accuracy mutation DNA detection technology to detect lung cancer EGFR L858R and 19Del

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