A comparison of diagnostic methods for the detection of bovine respiratory syncytial virus in experimental clinical specimens.
Virus shedding was monitored in nasal secretions of 12 calves experimentally infected with bovine respiratory syncytial virus (BRSV) using an antigen capture enzyme-linked immunosorbent assay (ELISA) detecting the nucleoprotein (NP) antigen of BRSV, by a polymerase chain reaction (PCR) amplifying the fusion protein of BRSV, and by a microisolation assay combined with immunoperoxidase staining for the F protein of BRSV. Under the conditions of this study, similar limits of detection and quantitative results were obtained from all three assays. BRSV was detected in nasal secretions of all calves for a minimum of 4 d. Virus shedding began on Day 2 after infection, peaked on Days 3-5, and was cleared in most calves by Day 8. The PCR, and to a lesser extent the ELISA, may detect virus shedding for a longer period after infection than virus isolation, possibly due to neutralization of the virus by rising mucosal antibody. Simulated environmental conditions likely to be experienced during transport of clinical field specimens markedly reduced the sensitivity of virus isolation but had a minimal effect on the results of the NP ELISA. Actual field transport conditions (overnight on ice) had minimal apparent effect on the results of the PCR assay. The less stringent specimen handling requirements, combined with low limits of detection, of both the nucleoprotein ELISA and PCR, indicate either of these assays are more suitable for diagnostic applications than virus isolation.
West
K
K
Department of Veterinary Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon. khw128@mail.usask.ca
Bogdan
J
J
Hamel
A
A
Nayar
G
G
Morley
P S
PS
Haines
D M
DM
Ellis
J A
JA
D000818:Animals; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D007124:Immunoenzyme Techniques; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis / Q000662:veterinary*; D018112:Respiratory Syncytial Virus, Bovine / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D017201:Virus Shedding