Immunofluorescence kit for detecting E-Cadherin expression of peripheral blood circulating tumor cells of patient with hepatocellular carcinoma and method

Abstract

The invention discloses an immunofluorescence kit for E-Cadherin expression on peripheral blood circulating tumor cells of a patient with hepatocellular carcinoma and a detection method. The immunofluorescence kit comprises a primary antibody suspension 100 mu l composed of 8% of PFA, 0.4% of PFA, 0.3% of TritonX-100, 10% of goat serum, rabbit anti-human E-Cadherin and rat anti-CD45, a secondary antibody suspension 100 mu l composed of fluorescently labeled goat anti-rabbit and fluorescently labeled goat anti-rat, and a DAPI tablet. The detection method comprises the following steps of: peripheral blood collection, peripheral blood sample pretreatment, sample separation, methanol fixation, film pasting, pap pen circle drawing, cell fixation, cell penetration, sealing, primary antibody incubation, secondary antibody incubation and nuclear staining, sealing and detection. According to the detection method provided by the invention, the E-Cadherin expression condition of the patient withadvanced or recurrent hepatocellular carcinoma can be detected without acquiring tissue specimens through needle biopsy, and real-time dynamic detection can be realized by utilizing a minimally invasive technology.

Description

technical field [0001] The invention provides an immunofluorescence kit and a detection method for detecting the expression of E-Cadherin in peripheral blood circulating tumor cells of patients with hepatocellular carcinoma, belonging to the technical field of molecular biology. Background technique [0002] Liver cancer is one of the common malignant tumors in my country, with a high mortality rate. It ranks third in the death order of malignant tumors after stomach and esophagus; it ranks second in rural areas in some areas, second only to gastric cancer. About 110,000 people die from liver cancer every year in my country, accounting for 45% of the world's liver cancer deaths. The average age of liver cancer is 44 years old. The pathological types of liver cancer are hepatocellular carcinoma, cholangiocarcinoma, and mixed liver cancer, among which hepatocellular carcinoma accounts for 70-80%, which is highly malignant and develops rapidly. If the treatment is not timely or...

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Problems solved by technology

[0008] Aiming at the disadvantages in the prior art of detecting patients with advanced or recurrent hepatocellular carcinoma that tissue samples cannot be obtained by real-time or repeated puncture, and thus cannot evaluate the real-time dynamic state of E-Cadherin in patients, and that existing detection methods are prone to false positives and false negatives, this paper The invention provides an immunofluorescence kit and detection method for detecting the expression of E-Cadherin in the peripheral blood of patients with hepatocellular carcinoma: the circulating tumor cells (CTC) in the peripheral blood of patients with advanced hepatocellular carcinoma are separated by using a membrane filtration device, and further Detection of E-cadherin expression in CTC by immunofluorescence technique

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