Detection of infectious bronchitis virus in chicken in Kerala by real time Taqman RT-PCR assay

Abstract

Chickens with signs of respiratory infection were screened for the presence of infectious bronchitis virus (IBV). Tissue samples were collected from dead and ailing chickens and were propagated intra-allantoically in embryonated chicken eggs and the allantoic fluid was harvested. The tissue samples and harvested allantoic fluid were probed with reverse transcriptase polymerase chain reaction (RT-PCR) with primers targeting 5’UTR of IBV. A total of 95 samples were tested, by RT-PCR. A Taqman probe labelled real time PCR assay targeting the 5’ UTR and Nucleocapsid (N) gene of IBV was standardised for the detection of IBV in all the 95 tissue and allantoic fluid samples to assess its efficiency. The real time PCR could detect IBV in all the 95 allantoic fluid and tissue samples including the samples, which were negative in preliminary detection. Hence, in the present study, the real time PCR assay was found to have equal efficacy on clinical samples and allantoic fluid and with a sensitivity of 100% and specificity of 90.90% in comparison with the conventional RT-PCR assay.

Keywords

• Embryonated chicken egg, Infectious bronchitis virus, N gene, Real time PCR, 5’UTR