Method for detecting pig plague virus specific antibody and its ELISA reagent kit

An enzyme-linked immunoassay reagent and swine fever virus technology, which is applied in the direction of measuring devices, material analysis and instruments through observation of the impact on chemical indicators, can solve the problem of affecting the effect of attenuated live vaccination, the vaccine is invalid, and cannot produce Virus immunity and other issues

Inactive Publication Date: 2008-03-19
TSINGHUA UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Maternal antibody content above a certain level and the existence of recessive infection will affect the effect of attenuated live vaccination, and may even make the vaccine ineffective, unable to produce immune protection against the virus, resulting in immune failure

Method used

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  • Method for detecting pig plague virus specific antibody and its ELISA reagent kit
  • Method for detecting pig plague virus specific antibody and its ELISA reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1, the preparation of the enzyme-linked immunosorbent assay kit that detects classical swine fever virus specific antibody

[0020] The enzyme-linked immunosorbent assay kit of detection classical swine fever virus specific antibody of the present invention comprises following reagent:

[0021] 1) Enzyme-labeled swine fever virus single epitope-specific antibody working solution;

[0022] 2) Hog fever virus antigen working solution for coating: a polypeptide solution containing an epitope whose amino acid residue sequence is the sequence shown in Sequence 1; 10 μg / ml, the solvent is coating buffer, that is, 0.1M, pH9.6 bicarbonate Sodium buffer;

[0023] 3) ELISA plate

[0024] 4) Positive serum control: hyperimmune serum of rabbits with classical swine fever virus;

[0025] 5) Negative serum control: serum of healthy rabbits not exposed to CSFV;

[0026] 6) Washing solution: PBS buffer containing 0.05% Tween-20 by volume (PBS buffer contains 5.54g / L Na ...

Embodiment 2

[0072] Embodiment 2, the detection effect experiment of kit of the present invention

[0073] The enzyme-linked immunosorbent assay kit prepared in Example 1 is used to detect the serum containing classical swine fever virus antibody, and the specific steps are as follows:

[0074] 1. Add the coated antigen working solution prepared in Example 1 to the microtiter plate, 50 μl per well, and overnight at 4°C;

[0075] 2. Pour off the coating solution, and wash the microtiter plate once with the washing solution;

[0076] 3. Block the microtiter plate with blocking solution for 30 minutes;

[0077] 4. Add 50 μl of positive serum control (primary swine fever virus rabbit hyperimmune serum) and negative serum control (no contact with CSFV) to each hole of the enzyme label plate after step 3 sealing. The healthy rabbit serum) and the swine fever virus-infected pig serum to be tested (purchased from the China Veterinary Drug Administration, including the original serum, 10-fold dil...

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Abstract

The invention discloses a method for detecting the specific antibody of classical swine fever virus and a special ELISA kit thereof. The kit includes a classical swine fever virus antigen and an enzyme-labeled classical swine fever virus single-epitope specific antibody; the said swine fever virus antigen is a polypeptide containing one or more than one amino acid residue sequence described in sequence 1. The detection reagent of the classical swine fever virus specific antibody of the present invention can carry out effective detection to the classical swine fever virus specific antibody by solid-phase antigen competition ELISA (blocking method); The B cell epitope ensures the differential diagnosis, and the high degree of conservation of the epitope among various strains ensures the specificity of detection.

Description

technical field [0001] The invention relates to a method for detecting the specific antibody of classical swine fever virus and a special ELISA reagent kit thereof. Background technique [0002] Swine fever is a highly contagious, highly pathogenic and highly lethal severe animal disease caused by swine fever virus, which is extremely harmful to the pig industry and is one of the legally reported severe infectious diseases determined by the World Organization for Animal Health , is also one of the first class of animal diseases identified in my country. For a long time, the measures taken by our country for the swine fever epidemic are "prevention and control". Prevention means vaccination with live attenuated swine fever vaccine (rabbitized attenuated swine fever vaccine) through immunization; control means controlling the epidemic, monitoring, isolation, and even partial "culling". In the prevention and control of swine fever, it is very important to monitor and detect th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N21/78
Inventor 陈应华戚昀王在时
Owner TSINGHUA UNIV
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