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Probe and kit for circulating tumor cell and neutrophil leucocyte identification

A technology for neutrophils and tumor cells, applied in the field of probes and kits for the identification of circulating tumor cells and neutrophils, can solve problems such as interference with CTC capture and staining, non-specific capture, cross-contamination, etc., to achieve Probability of Accurate Recurrence and Metastasis, Effect of Predicting Probability of Recurrence and Metastasis

Inactive Publication Date: 2016-08-24
广州迈景基因医学科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there are deficiencies in the identification of CTCs under the existing technical platform, mainly because neutrophils in large numbers in the blood are mistaken for CTCs. The reasons are: 1) The number of neutrophils is large, which is easily Non-specific capture; 2). Neutrophils express very weak CD45, and at the same time have non-specific transfection with CK; 3). The number of neutrophils is positively correlated with tumor progression, and in patients with metastatic lung cancer Medium, markedly increased peripheral blood neutrophils, interfering with CTC capture and staining

Method used

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  • Probe and kit for circulating tumor cell and neutrophil leucocyte identification
  • Probe and kit for circulating tumor cell and neutrophil leucocyte identification
  • Probe and kit for circulating tumor cell and neutrophil leucocyte identification

Examples

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Embodiment 1

[0051] A probe for identifying circulating tumor cells and neutrophils according to an embodiment of the present invention, which contains fluorescent dye A, CD45, cytokeratin and ANCA;

[0052] Among them, the fluorescent dye A is DAPI, and the cytokeratin is composed of Pan-CK, CK5 / 6, CK7 and CK20; both CD45 and ANCA are labeled with rhodamine (TRITC), and the cytokeratin is labeled with fluorescein isothiocyanate (FITC). ).

Embodiment 2

[0054] A kit for identifying circulating tumor cells and neutrophils according to an embodiment of the present invention, which contains the probe described in Example 1, and also contains cell washing solution, PBS and density gradient separation solution; wherein, cell washing The medium was RPMI medium containing 5% FBS.

[0055] Further, the kit also contains fixative, Triton X-100 and goat serum.

Embodiment 3

[0056] Example 3: Identification of Circulating Tumor Cells and Neutrophils by Immunofluorescent Staining

[0057] In this example, immunofluorescence staining was used to identify circulating tumor cells and neutrophils (the enrichment process of circulating tumor cells is as follows: figure 1 shown), the specific steps are as follows:

[0058] 1. Separation of peripheral blood mononuclear leukocytes / circulating tumor cells

[0059] 1) Blood sample collection: Inject 8 mL of peripheral venous blood into a disposable human venous blood sample collection container BD vacutainer to obtain a blood sample;

[0060] 2) Let the GSS warm to room temperature first, add 3mL GSS Centrifuge the tube at 1000×g for 30 seconds at room temperature, and throw the GSS under the porous filter membrane; dilute the blood sample 1:1 with PBS (8mL blood + 8mL PBS);

[0061] 3) Transfer 8 mL of diluted blood sample into a tube, transfer 16 mL of diluted blood sample into two tube, centrifuge...

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Abstract

The invention relates to a probe and a kit for circulating tumor cell and neutrophil leucocyte identification. The probe for circulating tumor cell and neutrophil leucocyte identification contains fluorescent dye A, CD45, cytokeratin and ANCA; the cytokeratin, the CD45 and the ANCA have fluorescent dye B with different colors. The kit for circulating tumor cell and neutrophil leucocyte identification comprises a probe. The fluorescent dye A, the CD45, the cytokeratin and the ANCA dyed CTC are used, so that the circulating tumor cells and the neutrophil leucocytes can be distinguished; the number of the circulating tumor cells in the peripheral blood of the lung cancer patient can be precisely identified; the reoccurrence and metastasis possibility of the lung cancer patient can be accurately predicted.

Description

technical field [0001] The invention relates to a probe and a kit, in particular to a probe and a kit for identifying circulating tumor cells and neutrophils. Background technique [0002] Lung cancer has become a common malignant tumor in all countries in the world. The "two firsts" of lung cancer are recognized at home and abroad, that is, "the first in incidence rate" and "the first in mortality". In the past 50 years, many countries have reported that the incidence and mortality of lung cancer have increased significantly. The incidence and mortality of lung cancer in men account for the first of all malignant tumors, and the incidence and mortality of lung cancer occupy the second place in women. It has become the malignant tumor that threatens human health and life the most. Recurrence and distant metastasis have become the main cause of death in lung cancer patients, and the occurrence of distant metastasis or recurrence in lung cancer patients can be evaluated early...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/533G01N1/38G01N1/30G01N21/64C09K11/06
CPCG01N33/6884C09K11/06C09K2211/1029C09K2211/1088G01N1/30G01N1/38G01N21/6486G01N33/533G01N2001/302G01N2800/12
Inventor 柯尊富王连唐杨冬成
Owner 广州迈景基因医学科技有限公司
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