Immunofluorescence kit for detecting C-MET expression of peripheral blood circulating tumor cells of hepatocellular carcinoma patient and detection method

A technology for 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 positives and false negatives of C-MET real-time dynamic status, etc., to reduce the loss of cells, Avoid edge effects, good stability

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 status of C-MET in patients, and that existing detection methods are prone to false positives and false negatives, this paper The invention provides

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  • Immunofluorescence kit for detecting C-MET expression of peripheral blood circulating tumor cells of hepatocellular carcinoma patient and detection method
  • Immunofluorescence kit for detecting C-MET expression of peripheral blood circulating tumor cells of hepatocellular carcinoma patient and detection method

Examples

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Embodiment 1

[0045] 1. Rinsing of 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 stand;

[0047] b) Add about 2mL of 75% medical alcohol into the filter with a Pasteur tube and wait for it to naturally filter out from the bottom of the filter;

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

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

[0050] Note: If the alcohol does not filter out naturally or the filter out is too slow during the filter rinsing process, please discard the filter immediately.

[0051] 2. Using immunofluorescence to detect the expression of C-MET in peripheral blood circulating tumor cells of patients with hepatocellular carcinoma:

[0052] (1) Peripheral...

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Abstract

The invention discloses an immunofluorescence detection method for C-MET expression on peripheral blood circulating tumor cells of a hepatocellular carcinoma patient. 100 [mu] l of a primary antibodysuspension composed of 0.4% of PFA, 0.3% of TritonX-100, 10% of goat serum, rabbit anti-human C-MET and rat anti-CD45, 100 [mu] l of a secondary antibody suspension composed of a fluorescently labeledgoat anti-rabbit and a fluorescently labeled goat anti-rat, and a DAPI mounting medium; the detection method comprises the following steps of: peripheral blood collection, peripheral blood sample pretreatment, sample separation, methanol fixation, film pasting, stroke circle grouping, cell fixation, cell penetration, sealing, primary antibody incubation, secondary antibody incubation and nuclearstaining, sealing and detection. According to the detection method provided by the invention, the C-MET expression condition of a patient with later-period or relapse 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.

Description

technical field [0001] The invention provides an immunofluorescence kit and a detection method for detecting the expression of C-MET 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, ranking third in the ranking of malignant tumor deaths after stomach and esophagus; in rural areas in some areas, it occupies the second place, 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 onset of liver cancer is 44 years old. Liver disease has a high degree of malignancy and develops rapidly. If the treatment is not timely or the treatment plan is improperly selected, the average survival time is half a year. Liver cancer may be asymptomatic in the earl...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/574G01N33/573G01N33/569
CPCG01N33/56966G01N33/573G01N33/57438G01N33/5748G01N33/582G01N2333/71G01N2333/912
Inventor 洪瑛侯景阳李文涛李琛琛
Owner 山东凯歌智能机器有限公司
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