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A Thymidine Kinase recombinant protein-based ELISA for detecting antibodies to Duck Plague Virus | Wen et al. Virology Journal 2010 7 77 http www.virologyj.eom content 7 1 77 VIROLOGY JOURNAL RESEARCH Open Access AThymidine Kinase recombinant protein-based ELISA for detecting antibodies to Duck Plague Virus Yongping Wen21 Anchun Cheng 21 1 2 3 Mingshu Wang 21 2 Han Ge21 Chanjuan Shen21 Sitong Liu1 Jun Xiang1 Renyong Jia2 Dekang Zhu2 Xiaoyue Chen1 3 Bei Lian1 Hua Chang1 and Yi Zhou2 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 .
