Double antibody sandwich ELISA kit used for Seneca viral antigen detection and application thereof

A technology of antigen detection and double-antibody sandwich, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complex operation steps and the inability to realize real-time detection of a large number of samples, and achieve the effect of avoiding cross-reaction and shortening the detection time

Inactive Publication Date: 2018-02-27
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, the detection methods used in the diagnosis of SVV have many procedures and complicated operation steps, and it takes at least one day for the detection to obtain the results. Therefore, it is impossible to realize the instant detection of a large number of samples in the event of an outbreak

Method used

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  • Double antibody sandwich ELISA kit used for Seneca viral antigen detection and application thereof
  • Double antibody sandwich ELISA kit used for Seneca viral antigen detection and application thereof
  • Double antibody sandwich ELISA kit used for Seneca viral antigen detection and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Example 1 Establishment of a double-antibody sandwich ELISA detection method for Seneca virus antigen detection

[0057] 1. Materials and methods

[0058] 1.1 Materials

[0059] 1.1.1 Experimental animals and antigens

[0060] 1.5-2.0 kg healthy male rabbits were purchased from the Lanzhou Veterinary Research Institute of the Chinese Academy of Agricultural Sciences; the inactivated antigen of porcine Seneca Valley virus (SVV) was prepared and provided by the Lanzhou Veterinary Research Institute of the Chinese Academy of Agricultural Sciences.

[0061] 1.1.2 Main reagents

[0062] 96-well microtiter plates were purchased from Costar Company, 2mol / L H 2 SO 4 Provided by the detection team of Lanzhou Veterinary Research Institute, fetal bovine serum was purchased from GIBICO, TMB was purchased from SurModics, carbonate buffer capsules were purchased from Sigma, and unstained protein markers were purchased from Fermentas.

[0063] 1.2 Method

[0064] 1.2.1 Preparati...

Embodiment 2

[0141] The assembling and using method of embodiment 2 kit

[0142] 1. Assembly of kit

[0143] (1) ELISA plate: Seneca virus guinea pig anti-IgG-coated ELISA plate, prepared according to the method in Example 1, and stored in a refrigerator at 4°C.

[0144] (2) 10× washing solution: the 10× washing solution contains 5v / v% Tween-20, 8w / w% sodium chloride, 0.2w / w% potassium chloride, 2.9w / w% disodium hydrogen phosphate and 0.2w / w% potassium dihydrogen phosphate aqueous solution, pH7.4;

[0145] (3) 10× sample diluent: the 10× sample diluent contains 5w / w% bovine serum albumin, 5v / v% Tween-20, 8w / w% sodium chloride, 0.2w / w% chloride An aqueous solution of potassium, 2.9w / w% disodium hydrogen phosphate and 0.2w / w% potassium dihydrogen phosphate, pH7.4;

[0146] (4) Enzyme-labeled antibody diluent: 0.01mol / L PBS buffer containing 1v / v% glycerol, 0.5w / v%BSA, 1w / v% casein and 0.05w / v%Procline300, pH7.4, according to Example 1 preparation.

[0147] (5) Enzyme-labeled antibody wo...

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Abstract

The invention discloses a double antibody sandwich ELISA kit used for Seneca viral antigen detection and application thereof. The kit comprises a Seneca viral guinea pig anti-IgG enveloped elisa plateand an HRP marked Seneca viral rabbit anti-IgG. The HRP marked rabbit anti-IgG is used for replacing primary antibodies and secondary antibodies in traditional ELISA, so that the operating steps aresimplified. Simultaneously, the SVV guinea pig anti-IgG is enveloped to the surface of a solid-phase carrier by changing an enveloping stabilization technology, the preparation technology of an enzyme-labeled antibody diluent is changed, an enzyme-labeled antibody working solution can be stored stably without changing the activity and valence, and a double antibody sandwich ELISA kit for SVV antigen specific detection and a detection method thereof are established. The vacancy of making up SVV ELISA antigen detection is proposed, the problem that existing virus separation and detection for SVVantigens are low in repeatability and sensitivity and complex in operational program is overcome, and effective technical means is provided for SVV antigen detection.

Description

technical field [0001] The invention relates to a virus detection kit and its application, in particular to a double-antibody sandwich ELISA kit for Seneca virus antigen detection based on Seneca virus (Seneca Valley virus, SVV) specific enzyme-labeled antibody technology and its application. The invention belongs to the technical field of biological detection. Background technique [0002] SVV, also known as Seneca Valley virus (SVV), is no stranger to U.S. swine herds. Over the past 30 years, nearly 20 cases of SVV infection have been identified. In the United States, SVV is commonly associated with idiopathic swine vesicular disease. SVA has been reported worldwide, including Canada, Australia, Italy, New Zealand, and Brazil. SVV is a non-enveloped, single-stranded RNA virus belonging to the picornaviridae family like porcine foot-and-mouth disease (FMD) and porcine vesicular virus. It may be the causative agent of porcine idiopathic vesicular disease, but this has n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/535
CPCG01N33/535G01N33/56983G01N2333/08
Inventor 郑海学田宏杨帆石正旺刘华南张克山朱紫祥李丹刘永杰何继军郭建宏蒋韬刘湘涛
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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