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.
Department of Comparative Medicine, College of Veterinary Medicine, University of Tennessee, Knoxville, Tennessee 37996, USA.
Nasser Z
NZ
Eleraky
Stephen A
SA
Kania
James F
JF
Evermann
Leon N D
LN
Potgieter
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