Immunofluorescence kit for detecting CEA 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 CEA gene mutation of peripheral blood circulating tumor cells of a non-small cell lung cancer patient and a detection method, and belongsto the technical field of molecular biology. The kit comprises 45mL of a diluent, 1mL of a destaining solution, 0.5mL of a staining solution A, 1mL 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 the detection method provided by the invention, the CEA 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 a 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 reagent kit and a detection method for detecting CEA gene mutation of 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 circula...

AI Technical Summary

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 status of CEA in patients, the present invention provides a non-diagnostic detection method for CEA 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 using a membrane filtration device, and the expression of CEA in CTCs is further detected by immunofluorescence technology

Images