A Thymidine Kinase recombinant protein-based ELISA for detecting antibodies to Duck Plague Virus

Author:

Wen Yongping,Cheng Anchun,Wang Mingshu,Ge Han,Shen Chanjuan,Liu Sitong,Xiang Jun,Jia Renyong,Zhu Dekang,Chen Xiaoyue,Lian Bei,Chang Hua,Zhou Yi

Abstract

Abstract Background Duck plague virus (DPV) is the causative agent of Duck Plague (DP) that causes significant morbidity and mortality throughout duck-producing areas of the world. The diagnosis of DP currently relies on the use of live or inactivated whole DPV virion as antigens in ELISA, but it is too laborious and expensive for routine application, and it is still difficult to get purified DPV virion with current technology. Results In this study, we describe the expression and purification of a recombinant Thymidine Kinase (TK) protein which makes antigen in an in-house developed, optimized and standardized ELISA. The specificity of the optimized TK-ELISA was evaluated by antisera against Duck Plague Virus (DPV), Duck Hepatitis B Virus (DHBV), Duck Hepatitis Virus (DHV), Riemerella Anatipestifer(R. A), Escherichia coli (E. coli) and Salmonella anatum (S. anatum). Only antisera against DPV yielded a specific and strong signal. In order to determine the sensitivity of the TK-ELISA, a panel of diluted sera was tested, and the minimum detection limit of 1:2560 (OD450 nm = 0.401) was obtained according to the endpoint cut-off (0.2438). The repeatability and reproducibility under the experimental conditions demonstrates a low variability (P > 0.05). The suspected sera samples (n = 30) were determined by TK-ELISA and the positive rate is 90% (27/30), and the TK-ELISA showed 83.33% (22+3/30) coincidence rate with the Serum Neutralization Test (SNT) and 90% (24+3/30) coincidence rate with the whole DPV virion based-ELISA (DPV-ELISA). When defining the dynamics of antibody response to attenuated live DPV vaccine, the maximum antibodies is reached after 4 weeks. Conclusions The results suggest that the TK-ELISA provides high specificity, sensitivity, repeatability and reproducibility for detection of anti-DPV antibodies in duck sera, and has the potential to be much simpler than DPV-ELISA and SNT for the sera epidemiological investigation.

Publisher

Springer Science and Business Media LLC

Subject

Infectious Diseases,Virology

Reference54 articles.

1. Baudet E: Een sterfte onder eenden in Nederland, veroorzaakt door een filtreerbaar virus (vogelpest). Tijdschr Diergeneesk 1923, 50: 455-459.

2. Fauquet CM, Mayo MA, Maniloff J, Desselberger U, Ball LA, (Eds): Virus Taxonomy: Eighth Report of the International Committee on Taxonomy of Viruses. California: Elsevier Academic Press; 2005.

3. Kaleta EF, Kuczka A, Kuhnhold A, Bunzenthal C, M BB, Hanka K, Redmann T, Yilmaz A: Outbreak of duck plague (duck herpesvirus enteritis) in numerous species of captive ducks and geese in temporal conjunction with enforced biosecurity (in-house keeping) due to the threat of avian influenza A virus of the subtype Asia H5N1. German Veterinary Journal 2007, 114: 3-11.

4. Weingarten M: Duck plague: clinical aspects, diagnosis, control. Tierarztliche Praxis 1989, 17: 53-56.

5. Leibovitz L, Hwang J: Duck plague on the American continent. Avian Dis 1968, 12: 361-378. 10.2307/1588237

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