Characteristic Differences in Reverse Transcription-Polymerase Chain Reaction Products of Ovine, Bovine, and Human Respiratory Syncytial Viruses

Abstract

Abstract. In reverse transcription-polymerase chain reactions (RT-PCR) and DNA hybridizations using primers and an oligonucleotide probe to the fusion (F) protein mRNA of bovine respiratory syncytial virus (BRSV), all the BRSV isolates and a goat isolate could be distinguished from prototype isolates of human respiratory syncytial viruses (HRSV) and ovine (sheep and bighorn sheep) respiratory syncytial viruses (RSV). However, RT-PCR amplifications with primers to sequences of the HRSV F protein mRNA resulted in amplified products of ≈ 243 bp if mRNA templates of subgroup A HRSV strains were present and slightly larger amplified products with subgroup B HRSV strains. No amplified products were observed in HRSV-primed RT-PCR with BRSV or goat or ovine RSV mRNA templates. Although the ovine RSV isolates were antigenically crossreactive with the goat RSV, HRSV and BRSV isolates, they were not amplified with either HRSV-or BRSVspecific primers in RT-PCR. These results confirm previous immunological comparisons suggesting that some ovine RSV isolates should be considered as distinct respiratory syncytial viruses.Bovine respiratory syncytial viruses (BRSV) are im-goats, 20 and Rocky Mountain bighorn sheep 28 ) have portant causes of acute respiratory disease in postwean-complicated this classification scheme. Early studies ing calves and feedlot cattle in the United States and that identified differences in the mean antibody titers Europe. 5,6,10,14,35 Human respiratory syncytial viruses of sheep infected with BRSV suggested that sheep might (HRSV) are the most important respiratory pathogens have their own related but distinct RSV. 27 Subsequent in infants and young children. 17,30 Both viruses are serologic studies showed that, although antigenically pneumoviruses within the family Paramyxoviridae and related to BRSV, some ovine RSV isolates had apare antigenically related; 8,16,21,25,29 however, important parent differences, including distinctive variations in antigenic differences are present, suggesting that each immunofluorescence staining patterns and serum neushould be considered a distinct respiratory syncytial virus (RSV). 7,21 Originally, HRSV was believed to be a monotypic virus, but reactivity of monoclonal antibodies to at least 4 different viral proteins have allowed its division into 2 major subgroups (A and B). 3,13,22 Recent studies have further divided the HRSV classification to identify 6 subdivisions within subgroup A, 3 subdivisions within subgroup B, and some HRSV isolates that cannot be classified in either. 2 Similarly, evidence now indicates that BRSV is distinct from HRSV, with at least 2 subgroups based on differences in the phosphoprotein and fusion (F) protein. 7 Virus isolates from other ruminant species (sheep, 12,18 tralizing titers. 1 Recent advances in RSV genetic research have demonstrated that a combined reverse transcription-polymerase chain reaction (RT-PCR) can be used to amplify and detect low levels of HRSV RNA from nasopharyngeal aspirates 26 and otitis media effus...

Evaluation of a multiplex immunoassay for bovine respiratory syncytial virus and bovine coronavirus antibodies in bulk tank milk against two indirect ELISAs using latent class analysis

“…For BRSV, a Norwegian study found that the current Norwegian strains of BRSV belonged to the same subgroup as other North European isolates, indicating that the within-country diversity is likely to be limited (Klem et al, 2014a). Additionally, cross-reaction is likely to be common, and has even been shown for isolates from different species (Oberst et al, 1993). Even though it seems unlikely that spatial antigenic diversity plays an important role as source of bias it cannot be excluded with complete certainty.…”

The Ovine Respiratory Syncytial Virus F Gene Sequence and its Diagnostic Application

“…15 PCR assays have been used in several studies for detection of bovine RSV. 16,21,32,33,35 In 1 study, specific bovine RSV primers amplified bovine RSV strains but did not amplify ovine RSV. The PCR assay used in a previous study was able to detect the bovine RSV but not the ovine RSV.…”

Comparison of Targeting F and G Protein Genes to Detect Bovine and Ovine Respiratory Syncytial Viruses

“…9,12,18,19 In the present study, the bovine RSVs tested (18 isolates from different sources) reacted identically and could be differentiated from ovine RSV in both PCR assays described in this study. Moreover, no PCR products were detected with human RSV, bovine parainfluenza virus 3, bovine herpesvirus 1, bovine virus diarrhea virus, bovine coronavirus, and bovine adenovirus in these 2 assays.…”

“…8 Polymerase chain reaction (PCR) assays have been recommended to detect bovine RSV infection in cattle. 9,12,18,19 Determining the contribution of bovine and ovine RSVs in respiratory tract disease in cattle and sheep is relevant to the complete understanding of RSV epidemiology and vaccine development in cattle and sheep. In a previous study, reverse transcriptase-PCR (RT-PCR) assay targeting F gene (RT-PCR F) 5 could detect and differentiate bovine and ovine RSVs.…”