Immunofluorescence kit for E-Cadherin expression of peripheral blood circulating tumor cells of patient with renal cell carcinoma, and detection method

Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to an immunofluorescence kit for detecting E-Cadherin expression of peripheral blood circulating tumor cellsof a patient with renal cell carcinoma, and a detection method. The immunofluorescence kit comprises a diluents 45 mL, decolorizing liquid 1 mL, a staining solution A 0.5 mL, a staining solution B 1mL, methanol 200 mu l, 2% PFA 200 mu l, 10% goat serum 100 mu l, a primary antibody suspension 100 mu l composed of mouse anti-CK, rat anti-CD45 and rabbit anti-E-Cadherin, a secondary antibody suspension 100 mu l composed of fluorescently-labeled goat anti-mouse, fluorescently-labeled goat anti-rat and fluorescently-labeled goat anti-rabbit, and a DAPI tablet. According to the detection method provided by the invention, the E-Cadherin expression condition of the patient with later-period or relapse renal cell carcinoma can be detected without acquiring a tissue specimen through needle biopsy.The immunofluorescence kit belongs to minimally invasive technology and can realize real-time detection.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to an immunofluorescence kit and a detection method for detecting the expression of E-Cadherin in peripheral blood circulating tumor cells of patients with renal cell carcinoma. Background technique [0002] Renal cell carcinoma (RCC) is one of the most common malignant tumors of the urinary system, its morbidity and mortality account for about 2-3% of systemic malignant tumors, and its incidence has been on the rise in recent decades. Radical surgery is an effective means for the treatment of renal cell carcinoma, but about 20% to 30% of renal cell carcinoma patients have developed distant metastasis at the time of diagnosis. Even after undergoing radical surgery for renal cell carcinoma, 20% to 40% of patients will still experience recurrence and metastasis. Previous studies have suggested that epithelial-mesenchymal transition (EMT) plays an important role ...

AI Technical Summary

Problems solved by technology

[0006] In order to overcome the inability of patients with advanced or recurrent renal cell carcinoma 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 non-diagnostic method for E-Cadherin gene mutation in peripheral blood circulating tumor cells of patients with renal cell carcinoma. Objective detection method: use a membrane filtration device to separate and obtain CTCs in the peripheral blood of patients with advanced or recurrent non-renal cell carcinoma whose tissue samples cannot be obtained, and further use immunofluorescence technology to detect the expression of E-Cadherin in CTCs

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