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Immunofluorescence kit for detecting E-Cadherin expression of peripheral blood circulating tumor cells of colorectal cancer patient

A technology for peripheral blood circulation and colorectal cancer, applied in the field of molecular biology, can solve the problems of false negatives, the inability of tissue samples to evaluate the real-time dynamic state of E-Cadherin in patients, and false positives of detection methods, so as to reduce the loss of cells and avoid the edge effect, good stability

Inactive Publication Date: 2020-11-20
山东凯歌智能机器有限公司
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Problems solved by technology

[0007] For the detection of patients with advanced tumors or recurrent colorectal cancer in the prior art, it is impossible to obtain tissue samples by real-time or repeated puncture, and thus cannot evaluate the real-time dynamic status of E-Cadherin in patients, and the shortcomings of existing detection methods are prone to false positives and false negatives, The present invention provides an immunofluorescence kit and detection method for the expression of E-Cadherin in the peripheral blood of patients with colorectal cancer, using a membrane filtration device to separate and obtain circulating tumor cells (CTCs) in the peripheral blood of patients with advanced colorectal cancer, and further Detection of E-cadherin expression on CTC by immunofluorescence technique

Method used

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  • Immunofluorescence kit for detecting E-Cadherin expression of peripheral blood circulating tumor cells of colorectal cancer patient
  • Immunofluorescence kit for detecting E-Cadherin expression of peripheral blood circulating tumor cells of colorectal cancer patient
  • Immunofluorescence kit for detecting E-Cadherin expression of peripheral blood circulating tumor cells of colorectal cancer patient

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Embodiment 1

[0049] 1. Use a membrane filtration device to separate and obtain CTCs in the peripheral blood of patients with advanced or recurrent colorectal cancer whose tissue samples cannot be obtained, and determine whether CTCs exist:

[0050] Collect 5ml of fasting blood from the median vein of the elbow for 8-12 hours, dilute the peripheral blood with 45ml of diluent, and then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes;

[0051] At fixed intervals, assemble the membrane filtration device: as attached figure 1 , figure 2 , image 3 As shown, the filtering device is composed of a filter 3, a filter membrane 7, a blood sample container 2, a waste liquid tank 5, and an iron stand 1;

[0052] Wet the filter 3 with 10ml of PBS, and then add the fixed peripheral blood sample into the blood sample container 2 of the membrane filtration device, so that it is naturally filtered by gravity, and the CTC is trapped on the filter membrane 7;

[0053] The dia...

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Abstract

The invention provides an immunofluorescence kit for E-Cadherin gene expression of peripheral blood circulating tumor cells of a colorectal cancer patient and a detection method, and belongs to the field of molecular biology. Particularly, the kit mainly comprises a primary antibody suspension composed of goat serum, mouse anti-CK, rat anti-CD45 and rabbit anti-E-Cadherin, and a secondary antibodysuspension composed of a fluorescently-labeled goat anti-mouse, a fluorescently-labeled goat anti-rat and a fluorescently-labeled goat anti-rabbit. The detection method mainly comprises the followingsteps of: collecting peripheral blood, treating the peripheral blood, filtering and enriching the circulating tumor cells, and detecting the E-Cadherin gene expression condition of the circulating tumor cells by an immunofluorescence method.

Description

technical field [0001] The invention provides an immunofluorescence kit and detection method for the expression of E-Cadherin in peripheral blood circulating tumor cells of colorectal cancer patients, belonging to the technical field of molecular biology. Background technique [0002] Colorectal cancer is one of the most common malignant tumors in the world, and its death rate ranked fourth among malignant tumors in 2018. Colorectal cancer is the third most common malignancy in the United States. In China, the epidemiological data of China for the whole year in 2018: the morbidity and mortality of colorectal cancer are both the fifth, with 376,300 new cases each year, and 25% of them have already metastasized when they are diagnosed. Colorectal cancer is mostly asymptomatic in the early stage, and it is mostly in the middle and late stage with metastasis when symptoms appear. Despite innovations in surgical and chemotherapy regimens, survival for patients with advanced col...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/574G01N33/569B01D29/05
CPCB01D29/05G01N33/56966G01N33/57419G01N33/57492G01N33/582G01N2333/705
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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