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Reagent and kit for staining quantitative detection of semen leucocyte subpopulations, and detection method thereof

A technology for quantitative detection of white blood cells, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inconvenience, unstable chromogenic substrates, and inability to detect, and achieve convenient use, suitable for batch detection, and easy operation Effect

Inactive Publication Date: 2006-04-05
深圳华康生物医学工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The leukocyte peroxidase method is one of the standard detection methods recommended by the WHO for leukospermia. out
Enzyme immunohistochemical staining is also recommended by WHO. This method uses the common antigen CD45 on the surface of leukocytes to detect leukocytes in semen. Since CD45 antigen is unique to leukocytes, this method is the most accurate method for detecting leukospermia. The existing enzyme Chromogenic substrates in immunohistochemical staining reagents are extremely unstable, and are ready-to-use, which is very inconvenient

Method used

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  • Reagent and kit for staining quantitative detection of semen leucocyte subpopulations, and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1. A reagent for quantitative detection of semen white blood cell population staining, comprising:

[0042] 1. Primary antibody: mouse anti-human CD45 IgG monoclonal antibody;

[0043] 2. Secondary antibody: IgG antibody of rabbit anti-mouse IgG;

[0044] 3. Enzyme conjugates: complexes of mouse IgG antigen and horseradish peroxidase (HRP)

[0045] 4. Substrate A: 3-amino-9-ethylcarbazole (AEC).

[0046] In the above reagents, the first antibody is used to bind to the CD45 antigen on the surface of leukocytes, the IgG-HRP complex is combined with the first antibody through the second antibody, and AEC is used to react with HRP to complete the final color development step. Therefore, as long as the antibody-providing animal in the first antibody is consistent with the animal to be opposed in the second antibody and the antigen-providing animal in the enzyme conjugate, it can be cattle, sheep, horses, etc.

Embodiment 2

[0047] Embodiment 2. A quantitative detection reagent for semen leukocyte population staining, comprising:

[0048] 1. First antibody freeze-dried powder: freeze-dried powder containing mouse anti-human CD45 IgG monoclonal antibody;

[0049] 2. Secondary antibody freeze-dried powder: freeze-dried powder of IgG antibody containing rabbit anti-mouse IgG;

[0050] 3. Enzyme conjugate solution: Tris-HCl buffer containing a complex of mouse IgG antigen and horseradish peroxidase;

[0051] 4. Substrate A solution: acetate buffer containing 4-6% of 3-amino-9-ethylcarbazole and 2-3% of dimethylformamide;

[0052] 5. Special slides (4 wells / block): slides coated with acid-resistant and hydrophobic coatings and printed with staining windows with a diameter of 0.8mm.

[0053] Compared with Example 1 in this example, the two antibodies are in the form of lyophilized powder, and the IgG-HRP complex and AEC are stored in corresponding buffers, which is more conducive to long-term storage ...

Embodiment 3

[0054] Embodiment 3. Another quantitative detection reagent for semen white blood cell population staining, including:

[0055] 1. First antibody freeze-dried powder: freeze-dried powder containing mouse anti-human CD45 IgG monoclonal antibody;

[0056] 2. Secondary antibody freeze-dried powder: freeze-dried powder of IgG antibody containing rabbit anti-mouse IgG;

[0057] 3. Enzyme conjugate: Tris-HCl buffer containing a complex of mouse IgG antigen and horseradish peroxidase;

[0058] 4. Substrate A: acetate buffer containing 4-6% of 3-amino-9-ethylcarbazole and 2-3% of dimethylformamide;

[0059]5. Special slides (4 wells / block): slides coated with acid-resistant and hydrophobic coatings and printed with staining windows with a diameter of 0.8mm.

[0060] Also includes:

[0061] 6. Fixative solution: a mixture containing 40-60% acetone, 40-60% methanol, and 3-8% 40% formaldehyde;

[0062] 7. Buffer: phosphate buffer containing 0.015% calcium chloride dihydrate, 0.02% po...

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Abstract

The invention discloses a semen white cell group coloring quantization testing agent, agent box and the testing method. The semen white cell group coloring quantization testing agent comprises: a first antibody: the IgG type monoclonal antibody of A type animal anti-human CD45; a second antibody: the IgG type antibody of B type animal anti-A type animal IgG; enzyme combiner: A type animal IgG type antibody and horse radish peroxides component; bottom object A: 3-aminoethane-9-ethyl carbazole. The agent can be made by the agent box type and can include special slide with acid-resistance water repellency paint and coloring window.

Description

【Technical field】 [0001] The invention relates to a reagent for in vitro diagnosis, a test kit and a detection method thereof, in particular to a quantitative detection reagent, a kit and a detection method for semen white blood cell group staining. 【Background technique】 [0002] Male reproductive tract infection is an important cause of male infertility, accounting for 4% to 10% of male infertility. In recent years, with the spread of sexually transmitted diseases and the increase of subclinical and recessive infections, the laboratory diagnosis of male reproductive tract infections is particularly important. Leukocytospermia refers to the density of white blood cells in semen exceeding 1×10 6 / ml. [0003] There are mainly two methods to detect leukocytes in semen by staining methods: leukocyte peroxidase method and enzyme immunohistochemical staining. The leukocyte peroxidase method is one of the standard detection methods recommended by the WHO for leukospermia. out...

Claims

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Application Information

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IPC IPC(8): G01N33/552G01N33/535
Inventor 傅剑华刘瑜胡家纯何林何小红
Owner 深圳华康生物医学工程有限公司
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