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Use of method for purifying, isolating and analyzing atypical circulating tumor cells and use of atypical circulating tumor cells

A tumor cell, purification and separation technology, applied in the field of atypical circulating tumor cells, which can solve problems such as omission

Pending Publication Date: 2020-10-27
CHANG GUNG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These circulating tumor cells undergoing EMT may reduce the expression of epithelial cell marker genes such as EpCAM and CKs. These circulating tumor cells that undergo EMT and are clinically highly associated with cancer metastasis may be missed

Method used

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  • Use of method for purifying, isolating and analyzing atypical circulating tumor cells and use of atypical circulating tumor cells
  • Use of method for purifying, isolating and analyzing atypical circulating tumor cells and use of atypical circulating tumor cells
  • Use of method for purifying, isolating and analyzing atypical circulating tumor cells and use of atypical circulating tumor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1. Whole blood sample processing and CD45-negative and EpCAM-negative cell counting methods

[0028] In this example, the experiments were approved by the Institutional Review Board of the Chang Gung Memorial Hospital. All blood sample donors obtained informed consent (approval number: 201601081B0), and all methods were carried out in accordance with relevant guidelines for clinical trials.

[0029] First, 4 mL of a whole blood sample derived from one individual is provided, and then blood cells are removed from the 4 mL of the whole blood sample. Use red blood cell lysate, 1L of red blood cell lysate contains 8.26g of NH 4 Cl, 1.19g of NaHCO 3 , 200 μL of 0.5M EDTA (ethylenediaminetetraacetic acid), pH 8, with a final pH of 7.3. Among them, the volume ratio of the whole blood sample to the red blood cell lysate is 1:10, and the reaction does not exceed 10 minutes, and the supernatant is removed by centrifugation after the reaction. Next, centrifuge at a rot...

Embodiment 2

[0032] Example 2. Specificity and Sensitivity Tests of CD45 Negative and EpCAM Negative Cells for Differentiation of Cancer Patients

[0033] In this example, the experiment was approved by the human experiment ethics department of Chang Gung Memorial Hospital. All blood sample donors obtained informed consent (approval number: 201601081B0), and all methods were carried out in accordance with relevant guidelines for clinical trials.

[0034] First, CD45-negative and EpCAM-negative atypical circulating tumor cells and their number information were obtained according to the operation procedure described in Example 1 above. Next, use the receiver operating characteristic curve (ROC curve) of the analysis population to set the cut-off value (cut-off value), where the cut-off value is the need to diagnose cancer, evaluate cancer prognosis, track and monitor cancer or evaluate cancer treatment effectiveness The average, median, or a value obtained by receiver operating characterist...

Embodiment 3

[0041] Example 3. The clinical significance of the number of typical circulating tumor cells and the number of CD45-negative and EpCAM-negative cells

[0042] In this example, the experiment was approved by the human experiment ethics department of Chang Gung Memorial Hospital. All blood sample donors obtained informed consent (approval number: 201601081B0), and all methods were carried out in accordance with relevant guidelines for clinical trials.

[0043] First, CD45-negative and EpCAM-negative atypical circulating tumor cells and their number information were obtained according to the operation procedure described in Example 1 above. Then, whether there is a difference in the average number of cells between healthy subjects and cancer patients by statistical analysis method, wherein the statistical method is Mann-Whitney U test (Mann-Whitney U test), P value less than 0.05 is considered Significant difference.

[0044] figure 2 Typical circulating tumor cells (A) and C...

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Abstract

The present invention relates to the field of detection, in particular to the use of a method for purifying, isolating and analyzing atypical circulating tumor cells and the use of atypical circulating tumor cells. The method for purifying, isolating and analyzing the atypical circulating tumor cells, as well as the atypical circulating tumor cells, can be used for preparing kits diagnosing cancer, assessing cancer prognosis, monitoring cancer, or assessing effectiveness of cancer treatment. The invention has the advantages of high throughput, high purity and high recovery rate, and no selection bias, successful purification, isolation and analysis of atypical circulating tumor cells. Through the analysis of atypical circulating tumor cells, cancer risk, cancer prognosis and effectivenesscan be assessed, and the sensitivity and accuracy of detection can be improved.

Description

technical field [0001] The invention relates to the field of detection, especially the application of the method for purifying, separating and analyzing atypical circulating tumor cells and the application of the atypical circulating tumor cells. Background technique [0002] Cancer metastasis is the main cause of death from cancer. Circulating tumor cells (CTCs), identified since 1869, are cells that escape from the primary tumor site into adjacent vasculature and subsequently reside in the blood circulation. There is already evidence that the presence of circulating tumor cells in the blood circulation is associated with cancer metastasis. Therefore, those skilled in the art have devoted themselves to studying circulating tumor cells to understand the mechanism of cancer metastasis. This research direction may stimulate those skilled in the art to develop new cancer treatment strategies. [0003] In addition, in terms of clinical application, the analysis of circulating...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/574
CPCG01N33/533G01N33/574G01N33/57484G01N2333/705G01N2800/52G01N1/30G01N33/57492G01N1/34G01N33/582C12N5/0694C12N5/0087
Inventor 吴旻宪廖家蓉洪凤君
Owner CHANG GUNG UNIVERSITY
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