Studying On Transcription And Expression Phase Of DPV TK Gene And The TK Based-ELISA

Following the indication and expression of TK gene, the transcription and expression phase of Duck plague virus (DPV) Thymidine Kinase (TK) Gene, the TK recombinant protein-based ELISA for detecting antibodies to Duck Plague Virus are performed in this paper. The results are as follows:1. Transcription and expression phase analysis of the DPV TK gene was carried by FQ-PCR and Western-blot. The transcription phase indicated that the starting of transcriptional is from 30min after infection of the DPV, following with a increase which appeared a highest peak at 4h. And then, with the decrease after 36h, it achieved lowest at 72h; The results of expression phase showed that the TK protein was firstly detected at 6h, followed a gradual increase and a highest peak at 8h. These results proved that the DPV TK gene possessed the characteristics of early time gene.2. The TK recombinant protein-based ELISA for detecting antibodies to Duck Plague Virus. In this study, we describe 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 Virus (DHV), Duck Hepatitis B Virus (DHBV), 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 (OD450nm= 0.401) was obtained according to the endpoint cut-off (0.2438). 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%coincidence rate with the Serum Neutralization Test (SNT) and 90%coincidence rate with the whole DPV virion based-ELISA (DPV-ELISA).