[Development of a rapid detection method for a virulent strain of duck enteritis virus based on real-time...

Duck enteritis virus (DEV) is a common herpesvirus that can cause acute, febrile, and septicemic infectious diseases in poultry, leading to tremendous economic losses in the poultry farming industry. Therefore, there is an urgent need for efficient and sensitive on-site rapid detection of DEV. Targeting the conserved fragment of the UL2 gene of DEV, specific recombinase polymerase amplification (RPA) primers and probes were designed. After optimization of the RPA reaction conditions, a fluorescent RPA isothermal rapid detection method for DEV was established. The results demonstrated that this detection method had the optimal reaction temperature of 40 ℃ and the optimal reaction time of 20 min, with a minimum detection limit of 2 fg/reaction for the virulent strain of DEV. The method specifically amplified only the virulent strain of DEV, showing no cross-reactivity with other common duck pathogens such as duck hepatitis virus and duck Tembusu virus. The established real-time fluorescent RPA method and qPCR were used to test 36 clinical samples of duck tissue. The RPA method demonstrated the sensitivity of 100%, specificity of 100%, and accuracy of 100%. The real-time fluorescent RPA detection method established in this study for the virulent strain of DEV features rapid detection, high sensitivity, and strong specificity, providing a feasible approach for the on-site rapid detection of duck plague.