Abstract
Context: Infectious bursal disease (IBD) is a major health threat to the world's poultry industry. This infectious outbreak in Morocco and many other countries throughout the world occurs despite intensive controls including proper biosafety practices and vaccination. The current research work was conducted to develop a rapid, sensitive, and specific TaqMan real-time reverse transcription-polymerase chain reaction (RT-PCR) method for rapid diagnosis of infectious bursal disease virus (IBDV). The intralaboratory validation of the conceptualized method was carried out with increased specificity, sensitivity, linearity, repeatability, and reproducibility. The diagnostic efficacy of the new assay was tested on 125 bursal samples collected from suspected cases distributed in different Moroccon regions and compared to the SYBR Green and conventional RT-PCR assay in terms of sensitivity and specificity. The new conceptualized method was more sensitive and less expensive compared to both conventional RT-PCR and Green method assays. The results obtained indicate that the standard curve exhibited a dynamic linear range (1 -1 x10(8) copies/, L) with a linear correlation (R2) of 0.999 between the cycle threshold (Ct) values vs. template concentration. The assay showed high sensitivity and ability in detecting 2X10(1) RNA copies per PCR reaction without any cross-reaction with other potential viruses belong avian type. The developed approach in the current research could be used to rapidly distinguish IBDV belongs to other pathogens. The developed test has been shown to be highly sensitive, specific, and reproducible and can be accurately used to detect avian IBDV RNA in clinical samples with high specificity within a short time compared to both conventional RT-PCR and Green method assays. (C) 2020 Friends Science Publishers