Porcine circovirus type 2 (PCV2) enzyme-linked immuno sorbent assay (ELISA) antibody detection kit

A technology for porcine circovirus and detection kits, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of difficult to popularize in grassroots units, long time consumption, and complicated test operations.

Inactive Publication Date: 2011-09-14
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are disadvantages such as complicated test operation, long time consumption, specific professional skills and equipment, expensive finished products, etc., and it is not easy to popularize in grassroots units.

Method used

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  • Porcine circovirus type 2 (PCV2) enzyme-linked immuno sorbent assay (ELISA) antibody detection kit
  • Porcine circovirus type 2 (PCV2) enzyme-linked immuno sorbent assay (ELISA) antibody detection kit
  • Porcine circovirus type 2 (PCV2) enzyme-linked immuno sorbent assay (ELISA) antibody detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1. The preparation method of porcine circovirus type 2 ELISA antibody detection kit

[0015] 1. Obtain PCV2-cap protein monoclonal antibody: PCV2-cap protein monoclonal antibody antibody is commercially available, and can also be prepared by conventional methods in the art. The method for preparing antibodies to known antigens is well known in the art; used in the present invention The PCV2-cap protein monoclonal antibody was purchased from RuralTechnologies, Incorporated;

[0016] 2. Obtain horseradish peroxidase (HRP)-mouse anti-pig IgG enzyme conjugate: mouse anti-pig IgG monoclonal antibody is commercially available, and can also be prepared by conventional methods in the art; the mouse used in the present invention Anti-pig IgG monoclonal antibody was purchased from Luoyang Servier Experimental Equipment Company; then it was labeled with the improved sodium periodate oxidation method; finally, the enzyme label was added with 1% bovine serum albumin, 1% case...

Embodiment 2

[0020] Embodiment two. The operation steps of porcine circovirus type 2 ELISA antibody detection kit:

[0021] 1. Dilute the sample to be tested 100 times with sample diluent, add 100ul to each well, add positive and negative control solutions at the same time, and set a blank control. Incubate at 37°C for 1 hour, wash the plate 5 times with washing solution, spin dry, add 100ul of enzyme-labeled antibody to each well, incubate at 37°C for 1 hour; wash the plate 5 times with washing solution, spin dry, add enzyme substrate solution, 100μl per well , develop color in the dark for 5-10 minutes, add stop solution, 50 μl per well. Use a microplate reader to measure the light absorbance at 450nm A 450nm .

[0022] 2. Result judgment: Cut off (CO value) = negative control light absorption value A 450nm ×2.1 times, sample value = sample light absorption value A 450nm / CO value, the sample value greater than 1 is judged positive; the sample value less than 1 and equal to 1 is judg...

Embodiment 3

[0023] Embodiment three: experimental results and analysis

[0024] 1. Specific detection: detect porcine PCV2 inactivated vaccine immune serum respectively with the test kit that the present invention makes, porcine PCV2 infects serum, porcine PCV1 infects serum, porcine blue ear virus antibody positive serum, classical swine fever virus antibody positive serum, pig pseudo Rabies virus positive serum and pig PCV2 negative serum were operated and judged according to the operation steps of the porcine circovirus type 2 ELISA antibody detection kit in the specific embodiment. The results showed that the pig PCV2 immune serum and infected serum were positive, and other samples The test was negative with a specificity of 100%.

[0025] 2. Sensitivity detection: detect the positive reference serum of different dilutions with the test kit that the present invention makes, operate and judge according to the operating steps of porcine circovirus type 2 ELISA antibody detection kit in ...

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Abstract

The invention relates to a porcine circovirus type 2 (PCV2) enzyme-linked immuno sorbent assay (ELISA) antibody detection kit. The detection kit contains a PCV2-Cap monoclonal antibody and PCV2-Cap pre-coated enzyme-labeled plate, blocking solution, sample diluent, an enzyme compound, washing concentrate, enzyme substrate solution and stop solution, wherein the nucleocapsid protein is purified Cap protein for forming virus-like particles. The specificity of the kit reaches 100 percent; and the sensitivity is 1:800. The kit is use for monitoring the porcine PCV2 antibody level, and definitely acquires the porcine immune level or infection state.

Description

technical field [0001] The invention relates to a detection kit, in particular to a porcine circovirus type 2 ELISA antibody detection kit. Background technique [0002] Porcine circovirus type 2 (PCV2) is the main pathogen that causes postweaning multisystemic wasting syndrome (PMWS) in weaned piglets. as a feature. In addition to causing PMWS, PCV2 is also associated with congenital tremor in piglets, porcine dermatitis nephritic syndrome, porcine respiratory syndrome (PDNS), and reproductive disorders in sows. Since the first discovery of PMWS in Canada in 1996, the associated diseases caused by PCV2 infection have been widespread all over the world. As my country's pig industry is increasingly threatened by PCV2 infection, there is an urgent clinical need to develop a PCV2 serum antibody detection kit. The PCV genome consists of a covalently closed circular single-stranded DNA, containing two large open reading frames ORF1 and ORF2, which encode the viral protein repl...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569
Inventor 朱薇刘汉平熊媛媛靖志强廖园园
Owner WUHAN CHOPPER BIOLOGY
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