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

A technology of peripheral blood circulation and tumor cells, applied in biological tests, measurement devices, material inspection products, etc., can solve the problems of inability to evaluate patients, false negatives, false positives of E-Cadherin real-time dynamic status, etc., to reduce the loss of cells, Good stability and avoid edge effects

Inactive Publication Date: 2020-09-18
山东凯歌智能机器有限公司
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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

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

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[0044] Example 1

[0045] 1. Rinse the filter of circulating tumor cell separator (YZYMED-A10)

[0046] a) Open the upper and lower plugs of the filter and place the filter on the rinse support;

[0047] b) Add about 2mL of 75% medical alcohol to the filter with a pasteurizer and let it naturally filter out from the lower mouth of the filter.

[0048] c) Add 8 mL of physiological saline to the filter for rinsing, remove the residual alcohol in the filter, and naturally filter out the physiological saline;

[0049] d) Repeat step 3 twice, leave a proper amount of physiological saline to ensure that the filter membrane is wet, and install the lower plug of the filter.

[0050] Note: If alcohol cannot be filtered out naturally or is filtered out too slowly during the filter rinse process, please discard the filter immediately.

[0051] 2. Using immunofluorescence to detect the expression of E-Cadherin in circulating tumor cells in the peripheral blood of patients with hepatocellular carcinom...

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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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Application Information

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IPC IPC(8): G01N33/58G01N33/574G01N33/569
CPCG01N33/56966G01N33/57438G01N33/57492G01N33/582G01N2333/705
Inventor 李琛琛洪瑛侯景阳李文涛
Owner 山东凯歌智能机器有限公司
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