Establishment and application of digoxigenin-labeled nucleic acid probe for detection of infectious bronchitis virus

According to the genomic sequences of N gene of infectious bronchitis virus(IBV) published in Genbank,a pair of primers was designed for amplifying the 582 bp fragment in RT-PCR experiments.The PCR product was labeled with digoxigenin as DNA probe for detection of IBV.The hybridization assay of specificity showed all RNA of IBV strains were positive,but other nucleotide extracted from NDV and PMV negative.The sensitivity test showed that as low as 10 pg of IBV’s RNA could be detected by DIG-labeled probe.Therefore,it was concluded that the DIG-labeled probe could be used for IBV detection.