Immunofluorescence kit and detection method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of small cell lung cancer patient

Abstract

The invention provides an immunofluorescence kit and a detection method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of a 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 PD-L1 expression condition of a patient suffering from advanced or recurrent 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 causedby an edge 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 PD-L1 gene mutation in peripheral blood circulating tumor cells of patients with 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. Small cell lung cancer (SCLC) accounts for about 15% to 20% of the incidence of lung cancer. Compared with non-small cell lung cancer, it has the characteristics of faster tumor doubling time, rapid growth and early metastasis. [0003] Circulating tumor cells (CTCs) are tumor cells shed from solid tumors and enter the peripheral blood circulation. Since their discovery in 1989, a variety of methods have been used to detect circulating tumor cells in peripheral blood. Recent studies have shown that its detection has import...

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Problems solved by technology

[0006] In order to overcome the shortcomings of patients with advanced or recurrent small cell lung cancer who cannot obtain tissue samples by real-time or repeated puncture, and thus cannot evaluate the patient's PD-L1 status, the present invention provides a non-diagnostic method for detecting PD-L1 gene mutations in peripheral blood circulating tumor cells of patients with small cell lung cancer. Objective detection method: use a membrane filtration device to separate CTCs in the peripheral blood of patients with advanced or recurrent small cell lung cancer whose tissue samples cannot be obtained, and further use immunofluorescence technology to detect the expression of PD-L1 in CTCs

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