Kit for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of prostate cancer patient and detection method

Abstract

The invention provides a kit for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of a prostate cancer patient and a detection method. The kit comprises a diluent, a destaining , a staining solution A, a staining solution B, an anti-human PD-L1 primary antibody, an enzyme-labeled goat anti-human secondary antibody, 0.1% of Triton X-100, 0.3% of H2O2 and a reagent A. The detection method comprises the following steps: separating and acquiring peripheral blood circulating tumor cells (CTC) by using a membrane filtration device, and detecting the PD-L1 expression condition of the peripheral blood CTCs by using an immunohistochemical technology. By utilizing the detection method provided by the invention, the PD-L1 expression condition of a patient with advanced or recurrent hepatocellular carcinoma can be detected without acquiring a tissue specimen through needle biopsy, and real-time dynamic detection can be realized by utilizing a minimally invasive technology;by adopting the separation device and the diluent, the circulating tumor cells are well separated, the interference of blood cells is avoided, the false positive result caused by the edge effect possibly generated in the dyeing process is avoided, the stability is good, the cell loss is reduced, and the detection accuracy is improved.

Description

technical field [0001] The invention provides a kit and a detection method for detecting PD-L1 gene mutation in peripheral blood circulating tumor cells of prostate cancer patients, belonging to the technical field of molecular biology. Background technique [0002] Worldwide, the incidence of prostate cancer ranks second among male malignancies, and cancer-specific death ranks sixth. According to the 2015 China Cancer Registration Annual Report, the incidence of prostate cancer in China ranked 9th among malignant tumors in 2011, and the incidence and mortality of prostate cancer in urban men ranked 6th and 9th among male malignant tumors, respectively. The difficulty in the treatment of prostate cancer is metastatic prostate cancer, especially castration resistance after endocrine therapy. At present, the treatment methods for metastatic prostate cancer after castration resistance are limited, and all conventional treatment methods (radiotherapy, chemotherapy, endocrine th...

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Problems solved by technology

[0007] Aiming at the disadvantages of detecting patients with advanced tumor or recurrent prostate cancer in real-time or repeated puncture to obtain tissue samples in the prior art, and thus not being able to evaluate the real-time dynamic status of PD-L1 in patients, and the existing detection methods are prone to false positives and false negatives, this paper The invention provides a kit and detection method for detecting PD-L1 gene mutations in peripheral blood circulating tumor cells of patients with prostate cancer: the circulating tumor cells (CTCs) in the peripheral blood of patients with advanced prostate cancer are separated by using a membrane filtration device, and the immune group is further used to Detection of PD-L1 expression on CTC by chemical technology

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