Immunofluorescence kit for detecting EGFR gene mutation of peripheral blood circulating tumor cells of non-small cell lung cancer patient and detection method

Abstract

The invention provides an immunofluorescence kit for detecting EGFR gene mutation of peripheral blood circulating tumor cells of a non-small cell lung cancer patient and a detection method, and belongs to the technical field of molecular biology. The kit comprises 45 mL of a diluent, 1 mL of a decolorizing solution, 0.5 mL of a staining solution A, 1 mL of a staining solution B, 200 [mu]l of methanol, 200 [mu]l of 2% PFA, 100 [mu]l of 10% goat serum, 100 [mu]l of a primary antibody working solution, 100 [mu]l of a secondary antibody working solution and a DAPI mounting medium. According to thedetection method provided by the invention, the EGFR expression condition of a patient suffering from advanced or recurrent non-small cell lung cancer can be detected without obtaining a tissue specimen through needle biopsy. The technology belongs to minimally invasive technology and can realize real-time detection. According to the method provided by the invention, a false positive result caused by an edge effect possibly generated in the dyeing process can be avoided, the stability is good, the cell loss is reduced, and the detection accuracy is improved.

Description

technical field [0001] The invention provides an immunofluorescence kit and a detection method for detecting EGFR gene mutation in peripheral blood circulating tumor cells of patients with non-small cell lung cancer, belonging to the technical field of molecular biology. Background technique [0002] Lung cancer is one of the main malignant tumors leading to the death of cancer patients. In my country, the morbidity and mortality of lung cancer rank first. Non-small cell lung cancer (NSCLC) accounts for about 85% of all lung cancers, and more than 70% of NSCLC patients are diagnosed at an advanced stage. Despite the continuous improvement of treatment techniques such as surgery, radiotherapy and chemotherapy, the 5-year survival rate of NSCLC patients is still less than 20%, and the main causes of death include local recurrence and distant metastasis. [0003] Circulating tumor cells (CTCs) are tumor cells shed from solid tumors and enter the peripheral blood circulation. S...

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Problems solved by technology

[0006] In order to overcome the shortcomings of patients with advanced or recurrent non-small cell lung cancer who cannot obtain tissue samples by real-time or repeated puncture, and thus cannot evaluate the patient's EGFR status, the present invention provides a non-diagnostic detection method for EGFR gene mutations in peripheral blood circulating tumor cells of patients with non-small cell lung cancer. Detection method: CTCs in the peripheral blood of patients with advanced or recurrent non-small cell lung cancer who cannot obtain tissue samples are separated by membrane filtration devices, and the expression of EGFR in CTCs is further detected by immunofluorescence technology

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