Isolation and characterization of mammalian orthoreoviruses using a cell line resistant to sapelovirus infection
- 1 November 2020
- journal article
- research article
- Published by Wiley in Transboundary and Emerging Diseases
- Vol. 67 (6), 2849-2859
- https://doi.org/10.1111/tbed.13655
Abstract
Porcine sapelovirus (PSV) is a causative agent of acute diarrhoea, pneumonia and reproductive disorders in swine. Since PSV infection interrupts the growth of other viruses due to its high replication capability in cell culture, the prevention of PSV replication is a keystone to the isolation of non-PSV agents from PSV-contaminated samples. In the present study, we established the PSV infection-resistant cell line N1380 and isolated three mammalian orthoreoviruses (MRV) strains, sR1521, sR1677 and sR1590, from swine in Taiwan. These Taiwanese isolates induced an extensive cytopathic effect in N1380 cells upon infection. The complete and empty virus particles were purified from the cell culture supernatants. Next-generation sequencing analyses revealed that the complete virus particles contained 10 segments, including 3 large (L1, L2 and L3), 3 medium (M1, M2 and M3) and 4 small (S1, S2, S3 and S4) segments. In contrast, the empty virus particles without genome were non-infectious. Phylogenetic analyses revealed that the Taiwanese strains belong to serotype 2 MRV (MRV2). We established an ELISA for the detection of IgG antibody against MRV2 by using the empty virus particles as the antigen. A total of 540 swine and 95 wild boar serum samples were collected in Japan, and the positive rates were 100% and 52.6%, respectively. These results demonstrated that MRV infection occurred frequently in both swine and wild boar in Japan. We established a cell line that is efficient for the isolation of MRV, and the ELISA based on the naturally occurring empty particles would be of great value for the surveillance of MRV-related diseases.This publication has 35 references indexed in Scilit:
- Complete Genome Sequence of a Porcine Orthoreovirus from Southern ChinaJournal of Virology, 2012
- Detection and molecular chracterization of porcine type 3 orthoreoviruses circulating in South KoreaVeterinary Microbiology, 2012
- Novel Human Reovirus Isolated from Children with Acute Necrotizing EncephalopathyEmerging Infectious Diseases, 2011
- Isolation and characterization of the first Chinese porcine sapelovirus strainArchiv für die gesamte Virusforschung, 2011
- A Post-entry Step in the Mammalian Orthoreovirus Replication Cycle Is a Determinant of Cell TropismOnline Journal of Public Health Informatics, 2010
- The diversity of the orthoreoviruses: Molecular taxonomy and phylogentic dividesInfection, Genetics and Evolution, 2009
- Avian reovirus: Structure and biologyVirus Research, 2007
- Extensive Syncytium Formation Mediated by the Reovirus FAST Proteins Triggers Apoptosis-Induced Membrane InstabilityJournal of Virology, 2005
- Putative Autocleavage of Reovirus μ1 Protein in Concert with Outer-capsid Disassembly and Activation for Membrane PermeabilizationJournal of Molecular Biology, 2005
- Sequence Characterization of Ndelle Virus Genome Segments 1, 5, 7, 8, and 10: Evidence for Reassignment to the Genus Orthoreovirus, Family ReoviridaeBiochemical and Biophysical Research Communications, 2001