Comparison of the Immunofluorescent Assay and Reverse Transcription-Polymerase Chain Reaction to Detect and Type Infectious Bronchitis Virus

Abstract

Indirect fluorescent antibody (IFA) assay and reverse transcription-polymerase chain reaction (RT-PCR) are two current methods commonly used for the detection of infectious bronchitis virus (IBV) and its serotypes. The objectives of this study were to compare the two methods relative to detecting IBV in chicken embryos that were artificially inoculated with Mass41, Ark99, or Mass41/Ark99 in serial embryo passages and in tracheas and cecal tonsils collected from vaccinated commercial flocks with and without respiratory problems. The tissues collected from these birds were used to inoculate chicken embryos. The chorioallantoic membranes collected from the inoculated embryos were examined by the IFA assay, and the allantoic fluids collected from the same embryos were subjected to the RT-PCR. The IFA assay and RT-PCR both were able to detect and type IBV when only one IBV strain was inoculated into embryos, regardless of the number of passages. When embryos were inoculated with Mass41 and Ark99 strains simultaneously, both strains were 100% detected by the RT-PCR. By the IFA assay, 20% of samples were positive for two strains and 80% were positive for the Ark only. Two of eight samples collected from healthy flocks that were vaccinated with Mass/Conn/Ark were positive by the IFA assay, and four of eight were positive by the RT-PCR. Sixteen percent (12/75) of the samples from birds experiencing respiratory problems were positive by the IFA assay and 35% (26/75) were positive by the RT-PCR. Results suggest that the RT-PCR is more sensitive than the IFA assay, especially when more than one strain of IBV is involved, but the IFA assay is rapid and cheaper than the RT-PCR.