Solid phase competition ELISA kit for detecting Seneca valley virus antibody, and applications thereof,

An antibody detection and solid-phase competition technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of complex operation steps, inability to realize real-time detection of a large number of samples, etc., to avoid cross-reaction and shorten the detection time. Effect

Inactive Publication Date: 2018-04-10
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 th

Method used

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  • Solid phase competition ELISA kit for detecting Seneca valley virus antibody, and applications thereof,
  • Solid phase competition ELISA kit for detecting Seneca valley virus antibody, and applications thereof,
  • Solid phase competition ELISA kit for detecting Seneca valley virus antibody, and applications thereof,

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Establishment of a solid-phase competitive ELISA detection method for Seneca virus antibody detection

[0053] 1. Materials and methods

[0054] 1.1 Materials

[0055] 1.1.1 Experimental animals and antigens

[0056] 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.

[0057] 1.1.2 Main reagents

[0058] 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.

[0059] 1.2 Method

[0060] 1.2.1 Preparati...

Embodiment 2

[0132] The assembling and using method of embodiment 2 kit

[0133] 1. Assembly of kit

[0134] (1) ELISA plate: an ELISA plate coated with Seneca virus inactivated antigen, prepared according to the method in Example 1, and stored in a refrigerator at 4°C.

[0135] (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;

[0136] (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, pH 7.4;

[0137] (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.

[0138] (5) Enzyme-labeled re...

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Abstract

The invention discloses a solid phase competition ELISA kit for detecting Seneca valley virus antibody, and applications thereof, wherein the kit comprises Seneca valley virus inactivated antigen-coated enzyme label plate and HRP-labeled Seneca valley virus rabbit anti-IgG. According to the present invention, the primary antibody and the secondary antibody in the traditional ELISA are replaced with the HRP-labeled rabbit anti-IgG so as to simplify the operation step; by changing the coating stabilization process, the surface of the solid phase carrier is coated with the SVV inactivated antigen, such that the enzyme-labeled antibody diluent preparation process is changed, the enzyme-labeled antibody working liquid can be stably stored without the change of the activity and the titer, and the solid phase competition ELISA kit for the specific detection of the SVV antibody, and the detection method thereof are established; and with the kit, the blank in the SVV ELISA antibody detection ismade up, the problems of low repeatability, low sensitivity and cumbersome operation procedures in the SVV antibody detection in the existing VNT detection are overcome, and the effective technical means is provided for the SVV antibody detection.

Description

technical field [0001] The invention relates to a virus detection kit and its application, in particular to the establishment of a SVV solid-phase competitive ELISA antibody detection kit based on 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 not been...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/532
CPCG01N33/532G01N33/56983G01N33/6854
Inventor 郑海学田宏杨帆石正旺刘华南张克山朱紫祥李丹曹伟军刘永杰
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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