Virucidal efficacy was evaluated for four recently available disinfectants: chlorine dioxide, potassium peroxymonosulfate, a quaternary ammonium compound, and citricidal (grapefruit extract). Sodium hypochlorite (3%) and tap water were used as positive and negative controls respectively. Feline herpesvirus, feline calicivirus, and feline parvovirus were exposed to the manufacturers' recommended dilutions of the evaluated disinfectants. Both chlorine dioxide and potassium peroxymonosulfate completely inactivated the three viruses used in this study. These disinfectants can aid in controlling nosocomial transmission of viruses with less of the deleterious effects of sodium hypochlorite. The quaternary ammonium compound evaluated in this study and citricidal were not effective against feline calicivirus and feline parvovirus.
Abstract. Ruminant respiratory syncytial viruses (RSVs) are classified into 2 subgroups, ovine RSV and bovine RSV. Although ovine RSV infects cattle, its contribution to bovine respiratory tract disease has not been established, which is an important issue for vaccine development in cattle. Diagnosis by virus isolation or serology has low or variable sensitivity and/or specificity and polymerase chain reaction (PCR) has been recommended as a rapid and sensitive technique for RSV detection. A simple procedure has been developed to detect and identify bovine and ovine RSVs. First, the nucleotide sequence of the ovine RSV fusion (F) gene was determined and compared with representative strains of bovine RSV and human RSV subgroups A and B. The ovine RSV F gene has 85 and 72-73% nucleotide identity with those of bovine RSV and human RSV, respectively. The predicted amino acid sequence of the ovine RSV F gene has 94 and 83-84% amino acid identity with those of bovine RSV and human RSV, respectively. Then PCR primers targeting a specific F gene fragment of bovine and ovine RSV were designed. The primers represented bases 85-103 and the complementary sequence to bases 510-493 of the ovine RSV F gene. A similar PCR product (426 bp) was obtained on agarose gel electrophoresis from bovine RSV and from ovine RSV. The products, however, were unique to the parent virus and could be distinguished by EcoRI or MspI restriction endonuclease cleavage. EcoRI cleaved the ovine product into 2 bands (285 and 141 bp) but failed to affect the bovine RSV PCR product. However, MspI cleaved the bovine product into 2 bands (229 and 197 bp) but had no effect on the ovine product. Also, this assay did not amplify any PCR product with human RSV. The reverse transcription-polymerase chain reaction (RT-PCR) followed by restriction enzyme digestion is a useful and practical approach for detection and differentiation of ruminant 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.
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