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Porcine pseudorabies virus gE IgM antibody colloidal gold immunochromatograohic assay test strip as well as preparation method and application thereof

A porcine pseudorabies virus and immunochromatographic detection technology, which is applied to measuring devices, analytical materials, biological material analysis, etc., can solve the problems of requiring special instruments and personnel, rarely detecting IgM antibodies, and long detection time, and achieves easy operation , The result is intuitive and easy to distinguish, and the effect is convenient to carry

Inactive Publication Date: 2014-08-13
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are many detection methods for this disease, mainly ELISA and PCR, etc., but these methods have disadvantages such as complicated operation, special equipment and personnel, and long detection time. Colloidal gold immunochromatography method is developed on the basis of immune diafiltration The rapid detection method, the method is simple to operate, good repeatability, high sensitivity, fast and intuitive results, and can be prepared in large quantities, simple process, low cost, suitable for large-scale on-site detection at the grassroots level
Moreover, at present, these methods mostly detect PRV antigen and IgG antibody, but rarely detect IgM antibody

Method used

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  • Porcine pseudorabies virus gE IgM antibody colloidal gold immunochromatograohic assay test strip as well as preparation method and application thereof
  • Porcine pseudorabies virus gE IgM antibody colloidal gold immunochromatograohic assay test strip as well as preparation method and application thereof
  • Porcine pseudorabies virus gE IgM antibody colloidal gold immunochromatograohic assay test strip as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Schematic diagram of the structure of the colloidal gold immunochromatographic detection test paper card for porcine pseudorabies virus gE IgM antibody 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 pseudorabies virus gE 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 5 The materials 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 anti-pig ...

Embodiment 2

[0058] Except that the amount of anti-pig IgM monoclonal antibody coated on detection line T 7 is 6 μg / cm; the amount of goat anti-mouse IgG coated on control line C 8 is 8 μg / cm; The labeling amount of the anti-porcine pseudorabies virus gE protein monoclonal antibody is 10 μg / mL, and the coating volume is 20 μL / cm; the method for colloidal gold-labeled anti-porcine pseudorabies virus gE protein monoclonal antibody: take the radius as 40nm pH value 8.2 Add 20 mL of colloidal gold with a concentration of 0.01%, add 200 μg of anti-porcine pseudorabies virus gE protein monoclonal antibody, combine it under stirring conditions, add BSA to a final concentration of 8%, and centrifuge to remove unbound anti-porcine pseudorabies virus gE protein monoclonal antibody and unstabilized colloidal gold particles and agglutinates, the deep red precipitate at the bottom of the centrifuge tube is dissolved with 2mL 0.01M pH 7.4 PBS buffer, which is the colloidal gold-labeled anti-porcine pseud...

Embodiment 3

[0060] The method of using the above-mentioned porcine pseudorabies virus gE IgM antibody colloidal gold immunochromatography test paper card:

[0061] Dilute 3-4 drops of the serum sample to be tested with 1mL 0.01M pH 7.4 PBS and drop 4-5 drops in the sample hole, and judge the result within 10-15 minutes. The red color of the control line C indicates that the test paper card is valid. The control line C and the detection line All Ts appear red, indicating that the sample PRV IgM antibody is positive, and only the control line C appears red, indicating that the sample PRV IgM antibody is negative.

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Abstract

The invention discloses a porcine pseudorabies virus gE IgM antibody colloidal gold immunochromatograohic assay test strip as well as a preparation method and application thereof. The test strip comprises a sample absorption region, a colloidal gold labeled probe region, a solidifying antibody region, a water absorption region, a bottom plate and a clamping shell, wherein the sample absorption region, the colloidal gold labeled probe region, the solidifying antibody region and the water absorption region are sequentially bonded on the bottom plate and are interlapped in the clamping shell; the sample absorption region is coated with purified porcine pseudorabies virus gE protein; the colloidal gold labeled probe region is coated with colloidal gold labeled porcine pseudorabies virus gE protein monoclonal antibody; the solidifying antibody region is sequentially provided with a detection line T coated with mouse anti-pig IgM monoclonal antibody and a control line C coated with goat anti-mouse IgG. The test strip is simple to operate, good in repeatability, high in sensitivity, rapid and intuitive in result, simple in process and low in cost, can be mass-prepared, and is suitable for mass field test and early diagnosis for porcine pseudorabies virus infection in swinery.

Description

[0001] technical field [0002] The invention relates to a detection test paper card and a preparation method thereof, in particular to a porcine pseudorabies virus gE IgM antibody colloidal gold immunochromatographic detection test paper card and a preparation method and application thereof. Background technique [0003] Porcine pseudorabies is a severe infectious disease caused by porcine pseudorabies virus (Pseudorabies Virus, PRV), characterized by reproductive impairment in pregnant sows, high fever and nervous stiffness in newborn piglets, and a high mortality rate of up to 100%. Because pigs are the most important storage hosts of the disease, both sick pigs and infected pigs are the source of infection of the disease, and the epidemic range is all over the world, seriously endangering the development of the world's pig industry. [0004] There are many detection methods for this disease, mainly ELISA and PCR, etc., but these methods have disadvantages such as complic...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/56983G01N33/531G01N33/558G01N2333/145
Inventor 牟林琳刘洁孙庆歌李建漆世华朱薇温文生谢红玲冯钊
Owner WUHAN CHOPPER BIOLOGY
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