Competitive ELISA detection method for DHAV-1 antibodies

A technology for duck hepatitis A virus and a detection method, which is applied in the field of ELISA detection of type 1 duck hepatitis A virus antibody competition, can solve the problems of difficulty in evaluating the titer of DHAV-1 egg yolk antibody, and achieve the effects of good stability and high specificity.

Active Publication Date: 2015-07-08
SHANDONG VOCATIONAL ANIMAL SCI & VETERINARY COLLEGE
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Problems solved by technology

[0023] Aiming at the deficiencies of the above-mentioned prior art, the present invention provides a type 1 duck hepatitis A virus antibody competition ELISA detection method, which solves the problem of DHAV-1 antibody titer and commercial DHAV-1 yolk antibody titer in breeding ducks and ducklings. Difficult to assess questions

Method used

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Examples

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Embodiment 1

[0043] This example provides a competitive ELISA detection method for initially establishing a DHAV-1 antibody using a monoclonal antibody.

[0044] ①The monoclonal antibody 3B2 was labeled with horseradish peroxidase (HRP). After the monoclonal antibody was purified by the conventional caprylic ammonium sulfate method, the purity of the monoclonal antibody was identified by SDS-PAGE. After the concentration of the monoclonal antibody was determined, the monoclonal antibody was labeled with horseradish peroxidase by the simple sodium periodate method.

[0045] ② Determine the coating concentration of antigen VP1 protein and the optimal dilution concentration of enzyme-labeled antibody by square array test, and determine the optimal dilution concentration of detection serum by calculating the inhibition rate of positive serum and negative serum. The established competitive ELISA method was used to detect the sample serum, and the critical value of the detection method was dete...

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Abstract

The invention provides a competitive ELISA detection method for DHAV-1 antibodies. The detection method solves the problem that at present, the valence of the DHAV-1 antibodies in the bodies of breed ducks and ducklings and the valence of commercial DHAV-1 egg yolk antibodies are difficult to evaluate. The detection method includes the steps of performing monoclonal antibody making, antigen coating, washing, closing, washing, enzyme-labeled antibody treating, washing and TMB developing; determining the OD450 nm value of each reaction hole; calculating the inhibition ratio of a serum sample, wherein the inhibition ratio (%)=(the OD value of an enzyme-labeled antibody-the OD value of the sample)*100%/the OD value of the enzyme-labeled antibody; if PI>=15.21%, judging that serum is positive; if PI<15.21%, judging that the serum is negative. According to the competitive ELISA detection method for the DHAV-1 antibodies, high specificity of monoclonal antibodies and high sensitivity of ELISA tests are achieved, stability of results is good, commercialization and automation can be achieved, and judgment can be made by naked-eye observation.

Description

technical field [0001] The invention relates to the technical field of animal medicine, in particular to a type 1 duck hepatitis A virus antibody competition ELISA detection method. Background technique [0002] Duck viral hepatitis (DVH) is an acute and highly fatal infectious disease of ducklings caused by duck hepatitis virus (DHV). It mainly affects ducklings aged 1 to 3 weeks. One of the main infectious diseases that endanger the duck industry. At present, the disease has occurred in various countries and regions that raise ducks all over the world. [0003] Duck viral hepatitis is caused by three different types of viruses, duck hepatitis virus type 1 (DHV-1), duck hepatitis virus type 2 (DHV-2) and duck hepatitis virus type 3 (DHV-3), among which Duck hepatitis virus types 2 and 3 are independent pathogens. In recent years, with the occurrence of duck viral hepatitis and the emergence of new strains, new standards have been established for the classification of duc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/577
Inventor 提金凤李志杰李汝春王海燕李舫
Owner SHANDONG VOCATIONAL ANIMAL SCI & VETERINARY COLLEGE
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