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Method for detecting mammaglobin and kit therefor

A technology of globulin and kit, which is applied in the field of mammoglobulin detection, can solve the problems of long qualitative detection time, small number of processed samples, and inability to quantitatively measure mammoglobulin. The results are accurate and reliable

Inactive Publication Date: 2006-05-03
初培国
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Therefore, one of the technical problems to be solved by the present invention is to provide a method for detecting mammoglobulin to overcome the technical defects of small number of processed samples, long qualitative detection time and inability to quantify mammoglobulin in the prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Collect 2.7-4.5ml of blood from the patient in a collection tube containing EDTA, mix well, remove the plasma, centrifuge the plasma for 15 minutes (2500×g), take the supernatant and freeze it. Latex particles (diameter 2 μm) were labeled with anti-mammaglobulin antibody RO28. Labeled latex particles were stored (2-8°C) in HEPES buffer (pH 8.2) containing 0.2 g / L sodium adida. Remove all reagents and let stand at room temperature for 10 minutes. Add 200 μl of PBS to each of the test tubes of normal healthy female serum negative control and mammoglobulin positive control, let stand for 10 minutes, and then mix well. Mix the gel particles in the test tube repeatedly. Use a slide with a test pool, first drop 20 μl of patient plasma, 20 μl of negative control, and 20 μl of positive control into different pools, shake them left and right for 1 to 2 minutes to mix well, then add one drop of latex particles each, and shake again for 2 to 2 minutes. The two were fully mixed ...

Embodiment 2

[0066] Latex particles (2 μm in diameter) were labeled with anti-mamiglobulin antibody RO48, and other steps were the same as in Example 1.

Embodiment 3

[0068] Double-sandwich enzyme-linked immunosorbent assay for detection of mammoglobulin, the steps are as follows:

[0069] 1) Dilute the specific anti-mammaglobulin RO48 antibody to 4 μg / ml. React 50 μl of the diluted RO48 antibody with a 96-well solid-phase carrier plate (200 ng / well), overnight at 4°C. Aspirate excess supernatant and wash away unbound antibodies and impurities;

[0070] 2) Add 250 μl skimmed milk (50 g / L, diluted with PBS buffer) and react at room temperature for 2 hours;

[0071] 3) Absorb excess supernatant and wash away unbound antibodies and impurities;

[0072] 4) Standard group: add 30 μl natural mamiglobulin to each well

[0073] 5) (diluted to 7 concentrations: 5ng / ml, 2.5ng / ml, 1.25ng / ml, 625pg / ml, 312pg / ml, 156pg / ml, 78pg / ml) and 10μl normal female serum;

[0074] Test sample group: add 30 μl diluent and 10 μl serum of breast cancer patient to be tested in each well;

[0075] Both the standard group and the test sample group have two duplicat...

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Abstract

The invention relates to method and agent box for detecting mammary gland globulin in the field of biology chemistry. The latex fixation leptospiral method comprises the steps of: a) mixing the mammary gland globulin antibody marked emulsion particle with the tested sample, b) quoting the test result by weather it has agglutination reaction, wherein the mammary gland globulin antibody marked emulsion particle is the emulsion particle which uses mammary gland globulin antibody R028 or R048 marked emulsion particle with the dilemma 2-10um; preserving the marked emulsion particle inside the HEPES buffer solution with pH 8.2. It also discloses a method for using the anti-mammary gland globulin monoclonal antibody and skin factor mark anti-mammary gland globulin monoclonal antibody to quantity test the mammary gland globulin by double clamp heart enzyme immune adsorption method. It also discloses a RT-PCR test mammary gland globulin expressing method and provides the agent box used to test the mammary gland globulin.

Description

technical field [0001] The invention relates to a method for detecting mammaglobulin and a kit thereof. It is a method for detecting mammoglobulin by latex agglutination inhibition method using latex particles labeled with anti-mammoglobulin monoclonal antibody. Background technique [0002] Breast cancer is a common malignant tumor in women, accounting for 22% of all female malignant tumors. Around the world, about 1.2 million women suffer from breast cancer every year, and 500,000 people die from breast cancer. In developed countries such as Western Europe and North America, the incidence of breast cancer ranks first among female malignant tumors. Breast cancer is twice as common in developed countries as it is in poor countries. According to statistics from 1992 to 1996, the incidence of breast cancer in the United States was 110.6 per 100,000 people. Although Asia is a region with a low incidence of breast cancer, the incidence of breast cancer in my country is also ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531G01N33/543G01N33/574
Inventor 初培国
Owner 初培国
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