Breast cancer circulating tumor cells detection system and kit

A tumor cell and detection system technology, applied in the field of breast cancer circulating tumor cell detection system and kits, can solve the problems of low specificity, false negative, wrong diagnosis, etc., and achieve the effect of high reliability of results and high separation purity

Active Publication Date: 2014-07-23
上海柏慧康生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because tumor cells are often in a state of dedifferentiation, tumor cells in the circulation are often highly heterogeneous, and some mutated tumor cells often escape capture due to the low specificity and low efficiency of the capture system, resulting in the phenomenon of slipping through the net. and get false negative results, leading to wrong diagnoses

Method used

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  • Breast cancer circulating tumor cells detection system and kit
  • Breast cancer circulating tumor cells detection system and kit
  • Breast cancer circulating tumor cells detection system and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The circulating tumor cell detection instrument listed in this embodiment includes a cell capture system, an analysis and counting system, and a waste bin. The analysis and counting system includes a fluorescence microscope and a high-throughput cell image analyzer. The circulating tumor cell detection instrument is also equipped with Computer interface, through the computer to control the analysis and counting system.

[0028] Referring to Fig. 1, the workflow of the circulating tumor cell detection instrument is as follows: an optimized cell protection agent is placed in the blood sample storage tube, and the blood sample is collected and stored by using the blood sample storage tube; The antibody has immunomagnetic microbeads made of homologous secondary antibody magnetic beads, and the sample blood in the blood sample storage tube is placed in the functional enrichment box of the cell capture system, and the immunomagnetic microbeads and the target cells in the sampl...

Embodiment 2

[0032] The use process of the functional enrichment box of the cell capture system

[0033] Open the functional enrichment package, the size is 10cm 3 , draw 7.5ml of peripheral blood sample with a syringe equipped with a sampling needle, anticoagulate with 3.8% sodium citrate 1 / 10 volume and centrifuge for 10min, 3000 rpm; draw the white blood cell layer in the middle, and put an equal volume of sterile double distilled water at room temperature Let stand for 5 minutes, centrifuge for 20 minutes; wash off the hemolyzed body in the supernatant, add an equal amount of normal saline, and centrifuge for 15 minutes; aspirate and discard the supernatant, add an equal amount of normal saline, centrifuge for 15 minutes, and save the obtained cells. Use a 1.0ml syringe to draw 0.5ml immunomagnetic microsphere sample, add it to a functional enrichment tube for incubation; after incubation for 30min, stain the cells, add phosphate buffer solution (pH=7.4, 0.1mol / L) to wash for 1-3 time...

Embodiment 3

[0035] Immunostaining analysis of tumor cells

[0036] Blank magnetic balls and immunomagnetic balls loaded with Epcam antibody were used to "capture" breast cancer tumor cells for immunostaining, and observed under a fluorescent microscope, as shown in image 3 with Figure 4 Shown:

[0037] It can be seen from the figure that the ability of the immune magnetic ball loaded with Epcam antibody to capture breast cancer cells is significantly higher than that of the blank magnetic ball.

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Abstract

The invention relates to a breast cancer circulating tumor cells detection system and a kit, the detection system comprises a breast cancer circulating tumor cells capture system, a magnetic sorting apparatus and an analysis counting system; the breast cancer circulating tumor cells capture system comprises a functional enrichment box, an immunization magnetic microballoon, a cell incubation apparatus and a fluorescence coloring agent; the automatic magnetic sorting apparatus is used for enriching the tumor cells and performing fluorescence dyeing; the analysis counting system includes a fluorescence microscope and a high flux cell image analyzer; and the kit comprises the immunization magnetic microballoon and the fluorescence coloring agent. According to the invention, a magnetic cell sorting technology and an immunocytochemistry technology are combined for detecting peripheral blood circulation tumor cells, analysis and counting are carried out on the tumor cells by the analysis counting system, and the analysis on the molecule and gene aspects of the circulating tumor cells can be realized with high efficiency.

Description

technical field [0001] The invention belongs to the field of breast cancer circulating tumor cell detection, in particular to a breast cancer circulating tumor cell detection system and a kit. Background technique [0002] Cancer is one of the major diseases that plague human life. Now the average annual cancer incidence rate is about 22%. Has basically spread, drugs are difficult to treat. The most important thing to beat cancer is early detection, most early cancers can be cured. Initially, the disordered growth of cancer cells made them lose their ability to adhere to each other. Loose tumor cells can enter blood vessels and lymphatic vessels, and enter blood circulation or lymph circulation. Cells that metastasize through blood circulation are called circulating tumor cells (circulating tumor cells). tumor cells; CTCs). In the early stage of tumor metastasis, circulating tumor cells (CTCs) can plant in distant organs along the circulatory system and gradually form met...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/532
CPCG01N33/5005G01N33/57415G01N2800/365
Inventor 沈鹤柏梁晓飞
Owner 上海柏慧康生物科技有限公司
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