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Method for recognizing peripheral blood circulating tumor cells of tumor patient based on isolation by size of epithelial tumor cells

A technology for peripheral blood circulation and tumor patients, applied in the field of molecular biology, can solve the problem of peripheral blood circulation tumor cells, and achieve the effect of improving specificity, accurate detection of CTC, and reducing false positive rate

Pending Publication Date: 2020-09-01
SHANDONG RES INST OF TUMOUR PREVENTION TREATMENT
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Problems solved by technology

In the prior art, there is no relevant record of identifying circulating tumor cells in the peripheral blood of tumor patients through cell size-based filtering (ISET)

Method used

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  • Method for recognizing peripheral blood circulating tumor cells of tumor patient based on isolation by size of epithelial tumor cells
  • Method for recognizing peripheral blood circulating tumor cells of tumor patient based on isolation by size of epithelial tumor cells
  • Method for recognizing peripheral blood circulating tumor cells of tumor patient based on isolation by size of epithelial tumor cells

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Embodiment Construction

[0044] Below in conjunction with accompanying drawing and embodiment the present invention is described as follows.

[0045] An example of using this technical method to separate, obtain and identify peripheral blood circulating tumor cells of 8 tumor patients (simultaneously detect 8 normal samples as negative controls).

[0046] 1. Using a membrane filtration device to separate and obtain CTCs in the peripheral blood of patients with advanced tumors

[0047]Collect 5ml of fasting blood for 8-12 hours from the median cubital vein, and use 45ml diluent (composition: 1mmol / L EDTA+0.1%BSA+0.1%trehalose+0.2%polyoxyethylene polyoxypropylene ether block copolymer) , the base solution is Tris-HCl buffer) to dilute peripheral blood, then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes;

[0048] 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 filte...

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Abstract

The invention relates to a method for recognizing peripheral blood circulating tumor cells of a tumor patient based on isolation by size of epithelial tumor cells (ISET), in particular for recognizingcirculating tumor cells which are captured by a physical method and are difficult to recognize by cell morphology, and belongs to the technical field of molecular biology. Whether CD45 and CD31 on suspected CTCs are expressed or not is detected by adopting a cell immunochemical method so as to exclude leukocytes and endothelial cells. Circulating tumor cells in peripheral blood of various cancerpatients can be relatively accurately recognized, and the technology belongs to fields of minimally invasive technologies and can be used for real-time dynamic detection. According to the method provided by the invention, CTC can be detected more accurately, the false positive rate of CTC detection is reduced, the specificity is improved, and the method has important clinical guidance significancein evaluating the illness state and the curative effect by clinically applying CTC detection.

Description

technical field [0001] The invention relates to a method for identifying circulating tumor cells in the peripheral blood of tumor patients based on cell size filtering (ISET), especially for circulating tumor cells that are captured by physical methods and difficult to identify by cell morphology, using CD45 and CD31 cell immunochemical methods to detect, It belongs to the technical field of molecular biology. Background technique [0002] The CTCBIOPSY® detection system separates CTCs based on cell size, and then stains the cells on the filter membrane by Wright-Giemsa staining. The cells detected by this method in strict accordance with the SOP operating requirements can be divided into the following categories: CTC, blood cells, heteromorphic cells, and nude-nuclear cells. However, the aberrant cells and suspected CTC cells found during the detection process are difficult to characterize using the CTC morphological judgment standard (Hoffman standard). Such cells cannot ...

Claims

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Application Information

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IPC IPC(8): G01N33/68C12N5/09C12M1/00C12M1/12
CPCG01N33/6872G01N33/56972C12N5/0694C12M23/48C12M29/04G01N2333/70589G01N2333/70503C12N2509/00C12N2509/10
Inventor 崔凯李胜李浩
Owner SHANDONG RES INST OF TUMOUR PREVENTION TREATMENT
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