Immunodiagnosis reagent kit for breast cancer and testing sheet

A diagnostic kit and immunohistochemical technology, applied in the field of immunology, can solve the problems of cumbersome operating procedures and high non-specificity

Active Publication Date: 2007-03-14
SHANGHAI ZHANGJIANG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently existing breast cancer diagnostic reagents generally have the problems of cumbersome operation procedures and high non-specificity, and cannot easily, quickly and accurately judge the expression of Her2 antigen in tumor tissue of patients

Method used

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  • Immunodiagnosis reagent kit for breast cancer and testing sheet
  • Immunodiagnosis reagent kit for breast cancer and testing sheet
  • Immunodiagnosis reagent kit for breast cancer and testing sheet

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Selection of tumor cells with different expression levels of Her2 antigen

[0040] Tumor cell selection was performed using the following steps:

[0041] 1) Alternative tumor cells include: human liver cancer cells QYC, human liver cancer cells KYC, human liver cancer cells LYX, mouse renal adenocarcinoma cells RENCA, D2F2 / E2 (the above five cells were obtained from the International Cooperation Tumor Research Institute of Second Military Medical University ), human epidermal cancer cell A431 (purchased from Shanghai Wuli Biotechnology Co., Ltd.) and MCF-7 (obtained from the National Cancer Institute of the United States), SK-BR-3 (purchased from Shanghai Wuli Biotechnology Co., Ltd.), HTB -20 (purchased from Guangzhou Wellman Company). After culturing in vitro to the logarithmic growth phase, various cells were taken for fluorescent staining;

[0042] 2) For cell counting, add the tumor cells to be tested into a special detection tube for flow cytometry, 2×...

Embodiment 2

[0049] Example 2 Preparation of Tumor Tissue Expressing Her2 Antigen Homogeneity

[0050] Preparation:

[0051] 1) Three tumor cells, D2F2 / E2, LYX and MCF-7, which were obtained in Example 1 and showed high, moderate and no expression of Her2 respectively, were selected and passaged in RPMI-1640 complete medium containing 10% fetal bovine serum Expansion, culture conditions are 5% CO 2 , 37°C and saturated humidity;

[0052] 2) For the three types of cells in the logarithmic growth phase, wash with PBS, centrifuge at 200g for 5 minutes, discard the supernatant, resuspend the cell pellet in PBS, count, and adjust the cell density to 1×10 7 / mL;

[0053] 3) The tumor cell suspension was inoculated subcutaneously in the neck of Balb / c nude mice respectively, and the local tumor growth and the general condition of the tested animals were regularly observed;

[0054] 4) When the tumor grows to 0.5cm 3 Left and right size (the formula for calculating tumor volume is x 2 y / 2, w...

Embodiment 3

[0061] Example 3 Preparation of Tumor Histopathological Sections and Detection of Surface Her2 Antigen Expression

[0062] 1. Production

[0063] 1) Preparation of paraformaldehyde phosphate buffer solution: 40g of paraformaldehyde, 500mL of 0.1mol / L PBS with pH7.4, mix the two and heat to 60°C, stir and add 1N NaOH dropwise until the solution is clear, then add PBS to The total volume is 1000 mL.

[0064] 2) After removing the tumor tissue, process the tissue as soon as possible, and try to keep the tissue fresh and not dry, and the tissue block should not be too large or too thick, especially the thickness should not exceed 1cm.

[0065] 3) The collected tissue is fixed in paraformaldehyde phosphate buffer solution. The amount of fixative solution should be sufficient, at least 20 times that of the tissue. Otherwise, the concentration of the fixative solution will decrease. Too long time may affect the dyeing effect.

[0066] 4) Dehydration, transparency, and wax-immersio...

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PUM

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Abstract

The disclosed testing block for Her2 express in mammary tissue includes: the Her2 high-express area, the express contrast area, the no-express contrast area, and the testing area for target tissue. This invention can provide accurate evaluation standard for clinic diagnosis on breast cancer.

Description

technical field [0001] The invention belongs to the field of immunology, and relates to a product for tumor diagnosis and detection. Further, the invention relates to a test piece for detecting the expression of Her2 in breast tissue and a kit containing the test piece. Background technique [0002] Breast cancer is a common malignant tumor in women, and its incidence rate ranks first among female malignant tumors in European and American countries. Although my country is an area with a relatively low incidence of breast cancer, the incidence rate is increasing year by year. The prognosis of most breast cancer patients with early surgery is better, but about 25-30% of the patients have overexpression of Her2 receptor in the cancer tissue, and the prognosis of these patients is significantly worse than that of patients with low expression of Her2, which is reflected in the recurrence rate after surgery The tumor-free survival rate and overall survival rate are lower than tho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/574
Inventor 郭亚军王皓侯盛钱卫珠
Owner SHANGHAI ZHANGJIANG BIOTECH
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