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Toxoplasmosis IgM antigen testing reagent and its application

A technology for antibody detection and toxoplasma gondii, which is applied to measurement devices, analyzes materials by chemical reaction, and analyzes materials by observing the impact on chemical indicators, which can solve the problems of long reaction time, insufficient sensitivity and complicated operation. and other problems, to achieve the effect of long response time

Active Publication Date: 2007-02-07
ZHUHAI S E Z HAITAI BIOLOGICAL PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are two kinds of capture method kits available at home and abroad, one is the antigen directly labeled with enzyme, but its sensitivity is not high enough; Concentration of Toxoplasma gondii antigen reacts with the specific antibody in the wells of the ELISA plate for a period of time, after washing the plate, add the enzyme-labeled toxoplasma monoclonal antibody of the working concentration to react in the plate wells for a period of time, the operation process is divided into three steps, the reaction It takes a long time and the operation is complicated. Users need to dilute Toxoplasma gondii antigen or enzyme-labeled Toxoplasma gondii mAb by themselves
Even in some kits, the anti-toxoplasma monoclonal antibody does not have a labeled enzyme, but needs to add an enzyme-labeled anti-mouse IgG antibody to complete the reaction

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Preparation of anti-toxoplasma monoclonal antibody

[0019] 1. Materials and methods

[0020] 1.1 BABL / c mice

[0021] SPF grade, female, 4-6 weeks old, weighing 18-22 grams.

[0022] 1.2 Toxoplasma gondii antigen for immunization

[0023] Toxoplasma gondii RH strains are preserved by our company. Toxoplasma gondii RH strain was subcultured and amplified in the peritoneal cavity of mice, and the protozoa were collected and washed with PBS, filtered through a G3 sand core funnel to remove impurities such as mouse peritoneal cells, and the parasites were collected. The purity of the protozoa was >98%. Repeated freezing and thawing 3 times, used as immunogen.

[0024] 1.3 Mouse myeloma cell lines

[0025] SP2 / 0 myeloma cell line.

[0026] 1.4 Other reagents and consumables are well known in the art and are omitted here.

[0027] 2. Immunization of mice

[0028] Take Toxoplasma gondii antigen plus an equivalent amount of 0.2% Lifosin, mix well, inject int...

Embodiment 2

[0055] Embodiment 2: the preparation of kit

[0056] The composition of this kit is:

[0057] (1) Monoclonal antibody human IgM microplate 96 wells (8) Concentrated washing solution (10 times) 50ml×1 bottle

[0058] (2) Enzyme marker 1 bottle (9) Sample diluent 50ml×2 bottles

[0059] (3) Positive control (direct use) 1 tube (10) Stop solution 1 bottle

[0060] (4) Negative control (direct use) 1 bottle (11) 1 sealed bag

[0061] (5) Critical control (direct use) 1 stick (12) Self-adhesive 2 pieces

[0062] (6) 1 bottle of chromogenic solution A (13) 1 copy of recording paper

[0063] (7) Chromogenic solution B 1 bottle (14) Instruction manual 1 copy

[0064] The 96-well monoclonal antibody human IgM microwell plate is coated with anti-human IgM-μ chain monoclonal antibody.

[0065] The enzyme marker is prepared by taking 2 self-made anti-toxoplasma monoclonal antibodies to mark horseradish peroxidase, mixing them in equal volumes, and then mixing them with toxoplasma an...

Embodiment 3

[0067] Embodiment 3: kit application:

[0068] 1. Sample requirements

[0069] In this test, serum or plasma is used for detection, and the dosage is 10 μl. Do not use bacteria-contaminated, lipemic or hemolyzed samples for serum or plasma. Collect serum according to the standard method, and store the sample at room temperature for no more than 8 hours. If the experiment is performed after 8 hours, the sample should be stored at 2-10°C, and if it is stored for more than 1 week, it should be stored at -20°C.

[0070] 2. Operating procedure

[0071] 1>. The kit should be equilibrated at room temperature for 20-30 minutes. Dilute the concentrated washing solution 10 times with distilled or deionized water.

[0072] 2>. Sample 1:100 dilution: Add 1ml of sample diluent into a clean small tube, add 10μl of the sample to be tested, and mix well.

[0073] 3>. Fix the slats on the plate frame, seal the remaining slats with self-adhesive and put them in a sealed bag for storage. A...

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PUM

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Abstract

The invention discloses an IgM antibody detection agent for toxoplasma, wherein it uses anti-human IgM-mu chain mono-clone antibody to pack the micro-pore plate, marking with the toxoplasma antigen and enzyme-label antigen for toxoplasma antigen and other assist agents to prepare the detection agent. This invention has high sensitivity and well specificity fit to convenient operation.

Description

technical field [0001] The invention relates to an antibody detection reagent, in particular to a detection reagent for toxoplasma gondii IgM antibody and its preparation method and use Background technique [0002] Existing methods for detecting IgM antibodies against Toxoplasma gondii are mainly enzyme-linked immunoassay and colloidal gold immunoassay, the basic theoretical basis of which is prepared by using the specific combination of antigen and antibody. Although the immunocolloidal gold method has the advantages of simple and fast operation, without the need for professionals and professional instruments, the specificity and sensitivity of the current immune colloidal gold method products at home and abroad are all worse than the enzyme-linked immunoassay; and the enzyme-linked immunoassay is also There are two kinds of indirect method and capture method. The operation of the indirect method kit is also simpler and faster, but the specificity and sensitivity are worse...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N21/78
Inventor 汤汉文
Owner ZHUHAI S E Z HAITAI BIOLOGICAL PHARMA
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