Detection of Newcastle disease virus in organs and faeces of experimentally infected chickens using RT-PCR

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Abstract
Newcastle disease (ND) is a highly contagious avian disease. Rapid diagnosis of ND plays an important role in controlling outbreaks. Until now, time-consuming isolation of ND virus (NDV) in embryonated chicken eggs was used for NDV detection. For rapid diagnosis, reverse transcription-polymerase chain reaction (RT-PCR) with RNA extracted from tissue samples and faeces originating from experimentally and contact-infected chickens was established. Conjunctiva, lung, caecal tonsil and kidney proved to be the most suitable organs. In infected animals, NDV was detected most frequently between day 4 and 6 post-infection. Contact-infected animals gave most positive results between day 6 and 13 after exposure. RT-PCR was also able to reproducibly detect NDV in faecal samples. The RT-PCR did not show any cross-reactivity with other avian paramyxovirus serotypes, and additionally offers the possibility of subsequent sequencing of the amplified DNA allowing pathotyping of the isolate.