Comparison of targeting F and G protein genes to detect bovine and ovine respiratory syncytial viruses.
In this study, 2 reverse transcriptase-polymerase chain reaction (RT-PCR) assays were developed and compared for simultaneous detection of bovine and ovine respiratory syncytial viruses (RSVs). One assay was based on a set of primers, which amplified a 426-bp fragment of either bovine or ovine RSV F gene (RT-PCR F). The F products could be distinguished by EcoRI or BstYI restriction endonuclease cleavage. In the other assay, a set of primers amplified a 542-bp fragment of either ovine or bovine RSV G gene (RT-PCR G). EcoO1091 and RsaI restriction enzymes were used to differentiate between the ovine and bovine PCR-G products. Sequencing of the PCR products confirmed the fidelity of both assays. The 2 assays were evaluated using 18 bovine RSV isolates, 1 ovine RSV, 1 bighorn sheep RSV isolate, 1 caprine RSV isolate, 2 human RSV isolates, and several other viruses associated with bovine respiratory tract disease. RT-PCR G may be more sensitive in detecting viral RNA. Because the target sequence of the F gene is more conserved than that of the G gene, RT-PCR F followed by the appropriate restriction enzyme cleavage may be superior to RT-PCR G to discriminate between the 2 ruminant RSV subgroups. This assay should prove useful for determining the relative contribution of ovine and bovine RSV to the pathogenesis of bovine respiratory tract disease.
Eleraky
Nasser Z
NZ
Department of Comparative Medicine, College of Veterinary Medicine, University of Tennessee, Knoxville, Tennessee 37996, USA.
Kania
Stephen A
SA
Evermann
James F
JF
Potgieter
Leon N D
LN
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000821:virology; D005814:Genes, Viral / Q000235:genetics*; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000662:veterinary* / Q000821:virology; D012136:Respiratory Syncytial Viruses / Q000235:genetics* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012757:Sheep Diseases / Q000175:diagnosis / Q000821:virology; D034561:Sheep, Domestic; D014759:Viral Envelope Proteins / Q000235:genetics
D014759:Viral Envelope Proteins