Development and evaluation of a real-time Taqman RT-PCR assay for the detection of infectious bronchitis virus from infected chickens.

摘要: It is important to rapidly differentiate infectious bronchitis virus (IBV) from disease agents like highly pathogenic avian influenza and exotic Newcastle virus, which can be extremely similar in the early stages of their pathogenesis. In this study, we report development testing a real-time RT-PCR assay using Taqman-labeled probe for rapid detection IBV. The amplifies 143-bp product 5'-UTR IBV genome has limit quantification 100 template copies per reaction. All 15 strains tested as well two Turkey coronavirus were amplified, whereas none other pathogens examined, positive. Evaluation was completed with 1329 tracheal swab samples. A total 680 samples collected antibody negative birds by assay. We 229 swabs submitted different diagnostic laboratories found 79.04% positive RT-PCR, only 27.51% isolation, reference standard test. also 120 at six time points experimentally infected dosages that, independent dose given, viral load trachea plateau 5 days post-inoculation. addition, an inverse relationship between given 14 post-inoculation observed. Finally, 300 samples, flock commercial broilers spray vaccinated field. percentage vaccine 3, 7, post-vaccination 58%, 65%, 83%, respectively, indicating that slightly more than half initially then subsequently transmitted flock. This observation significant because coronaviruses, have high mutation rate, revert pathogenicity when bird-to-bird transmission occurs. test described herein used distinguish respiratory pathogens, control virus. sensitive specific, quantitate genomic RNA clinical