Recombinant UL51 protein-based method for detecting duck plague virus antibody

A technology for duck plague virus and detection method, applied in the field of veterinary medicine, can solve the problems of time-consuming and laborious, hinder the large-scale application of ELISA method, and the sensitivity is not ideal, and achieve the effect of good specificity

Active Publication Date: 2011-04-06
SICHUAN AGRI UNIV
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Problems solved by technology

The classic method is the serum neutralization test, but its sensitivity is not ideal, and it is time-consuming and laborious, so it is not suitable for the detection of large quantities of serum samples
ELISA has the characteristics of strong specificity, high sensitivity, fast accuracy, and easy operation. It is an effective means for early rapid diagnosis of DPV infection and monitoring of immune antibodies; however, the complexity and purity of DPV whole virus purification methods are not ideal. Large-scale application of ELISA method (DPV-ELISA) coated with whole virus

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  • Recombinant UL51 protein-based method for detecting duck plague virus antibody
  • Recombinant UL51 protein-based method for detecting duck plague virus antibody
  • Recombinant UL51 protein-based method for detecting duck plague virus antibody

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Embodiment Construction

[0030] In order to make the object, technical solution and advantages of the present invention clearer, preferred embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings. In this example, the recombinant UL51 protein was firstly obtained by constructing the prokaryotic expression plasmid pET28a-UL51, transforming the expression bacteria, and fermenting and culturing, collecting a large amount of bacteria precipitates containing the recombinant UL51 protein, and then crushing the bacteria by ultrasonic waves, recombining UL51 protein inclusion body was obtained by washing, purification and gradient dialysis; at the same time, it was confirmed by Westernblotting analysis that the protein could have a strong immune reaction with anti-DPV positive serum; the coating concentration of the recombinant UL51 protein and the primary antibody, secondary Anti-dilution, thus preparing the duck plague virus antibody detection metho...

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Abstract

The invention relates to the field of animal medicine, in particular to a method for detecting duck plague virus antibody. The method particularly comprises the steps of the preparation of solid phase antigen, primary antibody combination, secondary antibody combination, developing, detection, judgment and the like. The method is an indirect enzyme-linked immuno sorbent assay (ELISA) method established based on purified recombinant UL51 protein, which has good specificity, and when duck hepatitis virus (DHV), Riemerella anatipestifer (RA), duck Escherichia coli (E.coli) are detected, the detection results are all positive; intra batch or batch-to-batch repeated tests show that variation coefficients are all less than 10%, and duck plague virus (DPV) vaccine immunized duck positive sera diluted at ratio of 1:3200 can be detected.

Description

technical field [0001] The invention relates to the field of animal medicine, in particular to a method for detecting duck plague virus antibody based on recombinant UL51 protein. Background technique [0002] Duck plague (duckplague, DP) is caused by DPV in the Herpesviridae family and is a highly fatal infectious disease common in waterfowl such as ducks, geese, and swans. It is directly related to the sustainable and stable development of the waterfowl breeding industry. Clinical and laboratory tests have confirmed that DPV attenuated vaccine is an effective biological agent for the prevention and control of DP, and the monitoring of DPV-specific antibodies is the key to evaluating the immune effect of DPV attenuated vaccine and formulating a reasonable immunization program. At present, the methods used to detect DPV antibodies mainly include neutralization test (neutralization test, NT), enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay, ELISA), agar ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569C12N15/38C12N15/70C07K14/03
Inventor 程安春汪铭书沈婵娟陈孝跃
Owner SICHUAN AGRI UNIV
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