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ELISA kit for identification of pig hoof-and-mouth disease immunity animal and taint animal

A technology for immunizing animals and swine foot-and-mouth disease, applied in the biological field, can solve problems such as unseen and unseen rapid identification and detection

Inactive Publication Date: 2008-07-23
ZHEJIANG UNIV
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Problems solved by technology

[0005] But so far, there is no ELISA detection method using the recombinant protein of the FMDV nonstructural protein 3ABC linear epitope and the mutant protein expression product of the structural protein VP1 antigen tandem epitope selected in this study as antigens for ELISA detection. An ELISA kit for simultaneous rapid identification and detection of porcine foot-and-mouth disease vaccine antibody and wild virus infection by structural and non-structural proteins

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  • ELISA kit for identification of pig hoof-and-mouth disease immunity animal and taint animal
  • ELISA kit for identification of pig hoof-and-mouth disease immunity animal and taint animal
  • ELISA kit for identification of pig hoof-and-mouth disease immunity animal and taint animal

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Embodiment Construction

[0024] The detection steps of the porcine foot-and-mouth disease ELISA rapid identification kit are as follows:

[0025] (1) Add 10 μL of the serum to be tested to 900 μL solution D for 100-fold dilution (take 20 μL for positive serum and negative serum, because they are 1:1 diluted in glycerol), dilute in Eppordorf tubes, and mix well ;

[0026] (2) Add 100 μL of the serum to be tested (including positive control and negative control) to the corresponding positions of the two 96-well plates, and place them in a wet box at 37°C for 1 hour;

[0027] (3) Add 200 μL solution G to each well for washing, 5 minutes each time, 6 times in total, in a micro shaker;

[0028] (4) Add 10 μL of solution C to 25 mL of solution D (2500-fold dilution), mix well, take 100 μL into each well of a 96-well plate, and incubate in a wet box at 37°C for 45 minutes;

[0029] (5) Repeat step 3;

[0030] (6) Add 100 μL of solution E to each well in the dark, and let it react in a humid chamber at 37°...

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Abstract

The invention relates to an ELISA test kit for identifying pig foot and mouth disease immunity animal and infected animal, wherein one orifice plate comprises the protein NS-AGD pure product of all linear antigen epitopes of FMDV non-structural protein 3ABC and another one orifice plate comprises the mutant protein CHA / 99 pure product of structural protein VP1 with wide antigen spectrum. The result is judged by cut-off that when in the infected animal non-structural protein test, the object sample OD492>=0.443, judges it positive, or else, judges it negative, when in the immunity animal structural protein test, when the object sample OD492>=0.450, judges it positive, or else, judges it negative. The test kit can quickly and synchronously identify pig foot and mouth disease vaccine antibody and wild virus infection, and can evaluate immunity effect according to the result.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to rapid diagnosis of animal epidemics and monitoring of vaccine immunity levels. A simultaneous detection technique was established to identify porcine Foot-and-Mouth Disease (FMD) vaccine-immunized animals and wild virus-infected animals by ELISA method, and the level of specific antibody in the serum after the vaccine immunization was determined. Background technique [0002] FMD is an acute, febrile and highly contagious infectious disease of cloven-hoofed animals caused by Foot-and-Mouth Disease Virus (FMDV). The disease is widely distributed around the world, spreads rapidly, and has a high incidence rate. The outbreak and prevalence of the disease not only directly endanger animal production, but also cause immeasurable losses caused by international trade barriers. There are 7 serotypes of foot-and-mouth disease virus, namely O, A, C, SAT1, SAT2, SAT3, and Asia1. There is no cros...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/68
Inventor 方维焕徐程江玲丽
Owner ZHEJIANG UNIV
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