Method for detecting specificity of breast cancer circulating tumor cells by using HER2 gene amplification method

A technique for tumor cell and gene amplification, applied in the field of molecular biology testing, to achieve the effect of protecting cells, increasing work difficulty, and good separation effect

Inactive Publication Date: 2019-05-21
GUIZHOU PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] With the increase in the mortality rate of female breast cancer year by year, breast cancer has become an important cause

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A specific method for detecting breast cancer circulating tumor cells by using a HER2 gene amplification method, comprising the following steps:

[0037]S1: Collect 8ml of peripheral blood from a patient with breast cancer, mix the collected blood sample with the lysate at a ratio of 1:5, heat up to 23°C after mixing evenly, add 2.5% glutaraldehyde by volume, and keep warm for 1 minute. Cool down to 15°C at a rate of 2°C, add 1.5% sodium glutamate drop by volume during the cooling period, let it stand for 2 minutes, wash and centrifuge with PBS to obtain breast cancer circulating tumor cells; the lysate is erythrocyte lysate , and contains dimethyl glycyrrhizinate which accounts for 0.5% by volume.

[0038] S2: Check the isolated breast cancer circulating tumor cells to check whether they exist, specifically:

[0039] (1) Centrifuge the sample of breast cancer circulating tumor cells to remove the supernatant, then add 12mL of 0.09mol / L KCl solution at 38.5°C, shake an...

Embodiment 2

[0052] A specific method for detecting breast cancer circulating tumor cells by using a HER2 gene amplification method, comprising the following steps:

[0053] S1: Collect 9ml of peripheral blood from a patient with breast cancer, mix the collected blood sample with the lysate at a ratio of 1:7, heat up to 25°C after mixing evenly, add 2.5% glutaraldehyde by volume, and keep warm for 1.5 minutes. Cool down to 16°C at a rate of 3°C, add 1.5% sodium glutamate drop by volume during the cooling period, let it stand for 2.5 minutes, wash and centrifuge with PBS to obtain breast cancer circulating tumor cells; the lysate is lysed red blood cells liquid containing 0.5% dimethyl glycyrrhizinate by volume.

[0054] S2: Check the isolated breast cancer circulating tumor cells to check whether they exist, specifically:

[0055] (1) Centrifuge the sample of breast cancer circulating tumor cells to remove the supernatant, then add 13mL of 0.09mol / L KCl solution at 38.5°C, shake and mix w...

Embodiment 3

[0068] A specific method for detecting breast cancer circulating tumor cells by using a HER2 gene amplification method, comprising the following steps:

[0069] S1: Collect 10ml of peripheral blood from a patient with breast cancer, mix the collected blood sample with the lysate at a ratio of 1:8, heat up to 28°C after mixing evenly, add 2.5% glutaraldehyde by volume, and keep warm for 2 minutes. Cool down to 17°C at a rate of 3°C, add 1.5% sodium glutamate drop by volume during the cooling period, let it stand for 3 minutes, wash and centrifuge with PBS to obtain breast cancer circulating tumor cells; the lysate is erythrocyte lysate , and contains dimethyl glycyrrhizinate which accounts for 0.5% by volume.

[0070] S2: Check the isolated breast cancer circulating tumor cells to check whether they exist, specifically:

[0071] (1) Centrifuge the sample of breast cancer circulating tumor cells to remove the supernatant, then add 15mL of 0.09mol / L KCl solution at 38.5°C, shake...

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Abstract

The invention discloses a method for detecting the specificity of breast cancer circulating tumor cells by using an HER2 gene amplification method. The method includes the following steps that: S1, the peripheral blood of a patient with breast cancer is collected, breast cancer circulating tumor cells are separated from the peripheral blood, so that the breast cancer circulating tumor cells can beobtained; S2, the separated breast cancer circulating tumor cells are tested, so that whether the breast cancer circulating tumor cells exit is checked; S3, the breast cancer circulating tumor cellsare cultured and amplified; S4, the cultured and amplified breast cancer circulating tumor cells are made into a test section; and S5, the HER2 expression of the breast cancer circulating tumor cellsis detected. The method of the invention has the advantages of simple operation and high detection accuracy. With the method adopted, false negatives and the like can be avoided, and the accuracy of the check of the breast cancer can be improved.

Description

technical field [0001] The invention relates to the technical field of molecular biology testing, in particular to a specific method for detecting breast cancer circulating tumor cells by using a HER2 gene amplification method. Background technique [0002] Malignant tumors will spread to other organs of the body through blood transmission, and tumor metastasis is the main cause of death of tumor patients. Tumor cells invade into the surrounding tissues of primary tumor cells, enter the blood and lymphatic vasculature, form circulating tumor cell CTCs, and transport to distant tissues, then exudate, adapt to the new microenvironment, and finally "seed" and "proliferate". ", "colonization", and the formation of metastases. Therefore, the early detection of CTC in the blood plays an important guiding role in the prognosis of patients, evaluation of curative effect and individualized treatment. [0003] HER2 is an important prognostic factor for breast cancer, and HER2 is als...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/574
Inventor 侯净魏娜朱海振戴民陈欢陆婧
Owner GUIZHOU PROVINCIAL PEOPLES HOSPITAL
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