Immunofluorescence kit and method for E-Cadherin mutation of peripheral blood circulating tumor cells of non-small cell lung cancer patient

Abstract

The invention provides an immunofluorescence kit and method for E-Cadherin mutation of peripheral blood circulating tumor cells of a non-small cell lung cancer patient, 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 the detection method provided by the invention, the E-Cadherin expression condition of the 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 anedge effect possibly generated in the dyeing process can be avoided, the stability is high, 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 E-Cadherin gene mutation in peripheral blood circulating tumor cells of patients with non-small cell lung cancer, and belongs 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 circul...

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Problems solved by technology

[0006] In order to overcome the inability of patients with advanced or recurrent non-small cell lung cancer to obtain tissue samples through real-time or repeated punctures, and to evaluate the status of E-Cadherin in patients, the present invention provides a method for E-Cadherin gene mutation in peripheral blood circulating tumor cells of patients with non-small cell lung cancer Detection method for non-diagnostic purposes: use a membrane filtration device to separate and obtain CTCs in the peripheral blood of patients with advanced or recurrent non-small cell lung cancer who cannot obtain tissue samples, and further use immunofluorescence technology to detect the expression of E-Cadherin in CTCs

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