Method for detecting E-Cadherin expression of colorectal cancer patient through peripheral blood circulating tumor cells

A technology for peripheral blood circulation and colorectal cancer, which is applied in separation methods, chemical instruments and methods, filtration separation, etc. effect of cell loss

Inactive Publication Date: 2020-09-08
山东凯歌智能机器有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In current clinical practice, the specimens for E-Cadherin detection in patients with colorectal cancer are mainl

Method used

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

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Experimental program
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Embodiment 1

[0046] 1. Use a membrane filtration device to separate and obtain CTCs in the peripheral blood of colorectal cancer patients who cannot obtain tissue samples to determine whether CTCs exist:

[0047] Collect 5ml of fasting blood for 8-12 hours from the median cubital vein, dilute the peripheral blood with 45ml of diluent, and then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes;

[0048] In a fixed interval, 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;

[0049] Wet the filter 3 with 10ml PBS, and then add the fixed peripheral blood sample to the blood sample container 2 of the membrane filtration device to make it filter naturally by gravity, and the CTC is trapped on the filter membrane 7;

[0050] The diameter of tumor cells is generally greater than 15 microns, and ...

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Abstract

The invention discloses a method for detecting E-Cadherin expression of colorectal cancer patient through peripheral blood circulating tumor cells. Specifically, a kit (45 mL of diluent, 1mL of decolorizing liquid, 0.5 mL of a staining solution A, 1 mL of a staining solution B, 100 [mu] L of a rabbit anti-human E-cad antibody, 100 [mu] L of goat anti-rabbit IgG/HRP, 100 [mu] L of 0.1% Triton X-100, 100 [mu] L of 0.3% H2O2, a reagent A, a reagent B, and 60 mL of a 6 * PBS buffer solution). The cancer period and recurrence condition of the patient are determined by collecting peripheral blood, pretreating the peripheral blood, enriching CTCs in the peripheral blood, dyeing and finally detecting the expression condition of genes by using an immunohistochemical technology.

Description

Technical field [0001] The present invention relates to a new method for detecting the expression of E-Cadherin, in particular to a method for detecting the expression of E-Cadherin in patients with advanced or recurrent colorectal cancer whose tissue samples cannot be obtained through peripheral blood detection. Background technique [0002] Colorectal cancer is one of the most common malignant tumors in the world, and the mortality rate in 2018 ranked fourth among malignant tumors. In the United States, colorectal cancer is the third most common malignant tumor. Domestically, China's annual epidemiological data in 2018: The incidence and mortality of colorectal cancer are both the fifth, with 376,300 new cases per year, and 25% have metastasized when they are diagnosed. Colorectal cancer is mostly asymptomatic in the early stage, and most of the symptoms appear in the middle and late stages with metastasis. Despite continuous innovations in surgical procedures and chemotherap...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/569B01D29/05
CPCB01D29/05G01N33/56966G01N33/57419G01N33/57492G01N2333/705
Inventor 侯景阳李文涛李琛琛洪瑛
Owner 山东凯歌智能机器有限公司
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