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Porcine circovirus type 2 (PCV2) IgM antibody colloidal gold immunochromatographic assay test paper, and preparation method and application thereof

An immunochromatographic detection, porcine circovirus technology, applied in biological testing, measuring devices, analytical materials, etc., can solve the problems of inability to distinguish specific PCV2IgM antibodies, requiring specialized instruments and personnel, and rarely detecting IgM antibodies, etc. Achieve intuitive and easy-to-recognize results, easy operation, and fast detection.

Inactive Publication Date: 2014-08-06
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The detection methods of this disease mainly include indirect immunofluorescence, PCR, etc. Although these methods can detect the level of PCV2 antigen or IgG antibody, they rarely detect IgM antibody, and the existing detection kits for detecting IgM antibody cannot distinguish specific PCV2 IgM antibody; and these The method has disadvantages such as complex operation, need for specialized instruments and personnel, and long detection time

Method used

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  • Porcine circovirus type 2 (PCV2) IgM antibody colloidal gold immunochromatographic assay test paper, and preparation method and application thereof
  • Porcine circovirus type 2 (PCV2) IgM antibody colloidal gold immunochromatographic assay test paper, and preparation method and application thereof
  • Porcine circovirus type 2 (PCV2) IgM antibody colloidal gold immunochromatographic assay test paper, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The schematic diagram of the structure of the porcine circovirus type 2 IgM antibody colloidal gold immunochromatographic detection test paper card is as follows figure 1 As shown, the horizontal surface of the test strip in the test paper card is sequentially divided into water absorption area 1, solidified antibody area 2, gold standard probe area 3, and sample absorption area 4 from left to right. After the test strip is loaded into the card case 6, it is the porcine circovirus type 2 IgM antibody colloidal gold immunochromatography test card; water absorption area 1, solidified antibody area 2, gold standard probe area 3, sample absorption area 4, bottom plate The materials of 5 are water-absorbing filter paper, nitrocellulose membrane, glass fiber membrane, polyester fiber membrane and polyethylene plate, and the cartridge 6 is a plastic cartridge with a viewing window and a sample hole. The detection line T 7 of the solidified antibody area 2 is coated with mouse ...

Embodiment 2

[0059] Except detection line T 7 coated with mouse anti-pig IgM monoclonal antibody amount of 3 μg / cm; control line C 8 coated with goat anti-mouse IgG amount of 7 μg / cm; gold standard probe region 3 coated with colloidal gold The labeling amount of the labeled anti-porcine circovirus type 2 Cap protein monoclonal antibody is 12 μg / mL, and the coating volume is 30 μL / cm; the method for colloidal gold-labeled anti-porcine circovirus type 2 Cap protein monoclonal antibody: take the radius 20mL of 40nm colloidal gold with a pH value of 9.2 and a concentration of 0.01%, added 240 μg of monoclonal antibody against porcine circovirus type 2 Cap protein, combined under stirring conditions, added BSA to a final concentration of 7%, centrifuged to remove untreated Combined anti-porcine circovirus type 2 Cap protein monoclonal antibody and unstabilized colloidal gold particles and agglutinates, and the deep red precipitate at the bottom of the centrifuge tube was dissolved with 2mL of 0....

Embodiment 3

[0061] The method of using the above-mentioned porcine circovirus type 2 IgM antibody colloidal gold immunochromatography test paper card:

[0062] Dilute 3-4 drops of the serum sample to be tested with 1mL 0.01M PBS with a pH value of 7.4, then drop 4-5 drops into the sample hole, and judge the result within 10-15 minutes. The red color of the control line C indicates that the test card is valid. Both the test line T and the test line T appear red, indicating that the sample IgM antibody is positive, and only the control line C appears red, indicating that the sample IgM antibody is negative.

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Abstract

The invention discloses a porcine circovirus type 2 (PCV2) IgM antibody colloidal gold immunochromatographic assay test paper, and a preparation method and application thereof. The test paper comprises a sample absorption region, a colloidal gold labeled probe region, a curing antibody region, a water absorption region, a backboard and a shell, wherein the sample absorption region, colloidal gold labeled probe region, curing antibody region and water absorption region are sequentially attached to the backboard and arranged in the shell in a superposition mode. A purified PCV2 Cap protein is coated on the sample absorption region; a colloidal gold labeled PCV2 Cap protein monoclonal antibody is coated on the colloidal gold labeled probe region; and the curing antibody region is sequentially provided with a detection line T coated mouse anti-pig IgM monoclonal antibody and a control line C coated goat anti-mouse IgG. The test paper is simple to operate, has the advantages of favorable repetitiveness, high sensitivity, quick and visual results, simple technique and low cost, can implement mass preparation, is suitable for mass field detection for basic level, and can be used for early quick diagnosis on swinery PCV infections.

Description

[0001] technical field [0002] The invention relates to a detection test paper card and a preparation method thereof, in particular to a porcine circovirus type 2 IgM antibody colloidal gold immunochromatographic detection test paper card and a preparation method and application thereof. Background technique [0003] Porcine circovirus type 2 (PCV2) infection can cause multisystemic wasting syndrome in weaned piglets, reproductive disorders in sows and other diseases. Since its discovery, it has widely existed all over the world, causing huge economic losses to the global pig industry. Therefore, the establishment of serological antibody detection method is of great significance to evaluate the immunity level and infection status of pig herds, and the detection of porcine circovirus type 2 IgM antibody provides a new diagnostic method for the early diagnosis of the disease. [0004] The open reading frame of PCV ORF2 encodes the viral nucleocapsid protein (Cap protein),...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6803G01N33/532G01N33/558G01N33/56911
Inventor 牟林琳刘洁朱薇李建漆世华温文生谢红玲冯钊
Owner WUHAN CHOPPER BIOLOGY
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