Colloidal gold immunochromatography test paper for antibodies of pseudorabies viruses of gE, gB and gD and preparation method

A porcine pseudorabies virus and immunochromatographic detection technology, which is applied in biological testing, measuring devices, analytical materials, etc., to achieve the effects of simple operation, good detection repeatability, and easy portability

Inactive Publication Date: 2014-05-14
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the reported PRV test strips can only detect antibodies against a single protein antigen (gE), and testers often

Method used

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  • Colloidal gold immunochromatography test paper for antibodies of pseudorabies viruses of gE, gB and gD and preparation method
  • Colloidal gold immunochromatography test paper for antibodies of pseudorabies viruses of gE, gB and gD and preparation method
  • Colloidal gold immunochromatography test paper for antibodies of pseudorabies viruses of gE, gB and gD and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Colloidal gold immunochromatography test paper planar structure area map of porcine pseudorabies virus gE, gB and gD antibodies figure 1As shown, the horizontal plane of the test paper is as follows from bottom to top: sample absorption area 1, gold label probe area 2, immobilized antigen and antibody area 3 and water absorption area 4, and immobilized antigen and antibody area 3 is coated with detection lines T1 31, test line T2 32, test line T3 33 and control line C 34.

[0034] Colloidal gold immunochromatographic test paper longitudinal section structure diagram of porcine pseudorabies virus gE, gB and gD antibodies (specific materials are used to represent each area) as shown in figure 2 As shown, the polyethylene plate 5 with a layer of polyvinyl chloride lining film is the support plate, the glass fiber membrane 6 is the sample absorption area, the polyester film 7 is the gold standard probe area, and the nitrocellulose membrane 8 is the immobilized antigen and...

Embodiment 2

[0053] Except that the coating amount of the detection line T1 is 5 μg; the coating amount of the detection line T2 is 5 μg, the coating amount of the detection line T3 is 5 μg, the coating amount of the control line C is 10 μg, and the gold standard probe colloidal gold-labeled mouse anti-pig IgG single The amount of anti-labeling is 20 μg of monoclonal antibody labeled with colloidal gold per mL, and the coating volume of gold-labeled probe is 5 μL; the method of colloidal gold-labeled porcine circovirus type 2 monoclonal antibody: take colloidal gold with a radius of 10nm and a concentration of 0.01% respectively 20mL and mouse anti-pig IgG monoclonal antibody 400μg, combined by magnetic stirring and shaking under the condition of pH 9.0, adding bovine serum albumin (BSA) and polyethylene glycol 20000 (PEG20000) as stabilizers, and making BSA The final mass concentration was 2%, and the final mass concentration of PEG20000 was 0.05%. The unbound mouse anti-pig IgG monoclonal...

Embodiment 3

[0055] Except that the coating amount of the detection line T1 is 2 μg; the coating amount of the detection line T2 is 2 μg, the coating amount of the detection line T3 is 2 μg, the coating amount of the control line C is 5 μg, and the gold standard probe colloidal gold-labeled mouse anti-pig IgG single The amount of anti-labeling is 10 μg of monoclonal antibody labeled with colloidal gold per mL, and the coating volume of gold-labeled probe is 8 μL; the method of colloidal gold-labeled porcine circovirus type 2 monoclonal antibody: take colloidal gold with a radius of 20nm and a concentration of 0.01% respectively 20mL and mouse anti-pig IgG monoclonal antibody 200μg, combined by magnetic stirring and shaking under the condition of pH 9.0, adding bovine serum albumin (BSA) and polyethylene glycol 20000 (PEG20000) as stabilizers, and making BSA The final mass concentration was 2%, and the final mass concentration of PEG20000 was 0.05%. The unbound mouse anti-pig IgG monoclonal ...

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Abstract

The invention discloses a colloidal gold immunochromatography test paper for antibodies of pseudorabies viruses of gE, gB and gD and a preparation method. The test paper comprises a sample absorption area, a gold mark probe area, an immobilization antigen and antibody area, a water absorption area and a supporting plate, and the sample absorption area, the gold mark probe area, the immobilization antigen and antibody area and the water absorption area are laid on the supporting plate and are partially overlapped in sequence. The gold mark probe area is covered with gold mark probes which are mouse anti-swine IgG antibodies marked by colloidal gold. The immobilization antigen and antibody area is provided with a detection line T1 covered with pseudorabies virus gE protein, a detection line T2 covered with pseudorabies virus gB protein, a detection line T3 covered with pseudorabies virus gD protein and a control line C covered with goat anti-mouse IgG antibodies. The test paper is fast in detection, high in accuracy, strong in specificity, easy and convenient to carry and operate and capable of being used for differential diagnosis on PRV wild poisonous infection and vaccine immunity and evaluation on the PRV vaccine immunity effect at the same time.

Description

[0001] technical field [0002] The invention relates to a detection test paper and a preparation method thereof, in particular to a colloidal gold immunochromatographic detection test paper for porcine pseudorabies virus gE, gB and gD antibodies and a preparation method. Background technique [0003] Pseudorabies is an acute infectious disease caused by pseudorabies virus (PRV), including a variety of domestic and wild animals. Pigs are the main storage host and source of infection for pseudorabies. Healthy pigs can be infected with the disease by direct contact with sick pigs and infected pigs. Adult pigs are often insidiously infected; infected pregnant sows have symptoms such as miscarriage, stillbirth, weak fetus, and mummified fetuses; infected newborn piglets have fever and neurological symptoms, and even collapse to death, with a mortality rate of up to 100%. [0004] Since pseudorabies was first discovered in the United States in 1902, the disease has sprea...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6803G01N33/532G01N33/558G01N33/56994G01N2333/032
Inventor 廖园园漆世华秦伟朱薇刘洁孙庆歌郑良益谢红玲温文生冯钊
Owner WUHAN CHOPPER BIOLOGY
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