Abstract:

A pair of primers was designed according to a conserved region of infectious bronchitis virus (IBV) N gene （FJ767920）, reverse transcriptase-polymerase chain reaction (RT-PCR) for rapid detection of IBV was developed by optimizing amplification conditions such as concentrations of primers for RT-PCR. A 318 bp fragment was amplified from IBV M41 strain and Nephropathogenic IBV HN99 strain, respectively, but not from infectious laryngotracheitis virus, Newcastle disease virus and Mareks disease virus. This amplified fragment was cloned into pGEM T-Easy vector and sequenced. The result showed this amplified fragment is specific for IBV cDNA. This method could detect the template cDNA of 0.6 pg for IBV. Clinical detection of IBV by RT-PCR showed that this improved RT-PCR approach could be a fast, simple and specific detection method for IBV.