Quantitative Study On Duck Plague Virus Infection And Alpha-interferon MRNA Level

Duck plague virus (DPV), an alphaherpesvirus, is the causative agent of duck plague (DP) that is an acute, contagious disease in ducks, goose, swans and other waterfowls. DP has been responsible for considerable economic losses to the commercial duck industry. For prevention and control of the disease, it is very important to detect virus rapidly and to clarify the process of infection, transmission and pathogenesis. Methods routinely used to identify the virus were troublesome and deficient in sensitivity and specificity, and not suitable for rapid diagnosis, large-scale quarantine and epidemic investigations. Therefore, it is urgently needed to develop a rapid and sensitive assay for DPV detection. While conventional PCR has been used to detect DPV, qualitative PCR is inability to quantify the virus DNA accurately.Real-time PCR, as a new method developed in recent years, provides a novel means to quantitatively analyze DNA and mRNA expression levels, and has proven to be very useful in virology researches such as rapid detection, virus pathogenesis, and host-virus interactions. In this study, following works focused on DPV infection were carried out based on real-time PCR assay.1. Development of a real-time PCR for DPV. Primers and probe were designed according to the sequence of DNA polymerase gene of DPV, and two real-time PCR assays were established based on TaqMan and SYBR Green I methods respectively. The results showed that both assays have a high sensitivity, specificity, repeatability and able to quantify the DPV DNA accurately. The detection limit was as low as 23 copies for TaqMan method and 230 copies for SYBR Green I method. The linear range was 2.3×...