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Isolation and propagation of the animal rotaviruses in MA-104 cells--30 years of practical experience.
- Source :
-
Journal of virological methods [J Virol Methods] 2015 Oct; Vol. 223, pp. 88-95. Date of Electronic Publication: 2015 Jul 31. - Publication Year :
- 2015
-
Abstract
- A total of 136 rotavirus positive samples from diarrhoeic animals of different species were submitted for isolation and cultural propagation of rotavirus on MA-104 cells. The samples were collected from animals with diarrhoea, between 1980 and 2010, originating from herds or farms located in several parts of Germany. Rotaviruses of species A were isolated from 102 faecal samples in cultures of MA-104 cells under the following conditions: pre-treatment of virus with trypsin, incorporation of trypsin into culture medium, use of roller cultures, and centrifugation of the samples on the cells. The cell culture adapted viruses produced a cytopathic effect, accompanied by the release of cells from the glass surface of the cultivation vessels. After 10 passages the virus isolates yielded titres between 10(5.5) and 10(7.5)ml(-1) TCID50. Isolation and serial propagation of the virus in MA-104 cells was confirmed by immunofluorescence assay, transmission electron microscopy, and polyacrylamide-gel electrophoresis of viral dsRNA. Eight (5.9%) of the electrophoretic profiles were characteristic of species B or D rotaviruses, which were not replicated in MA-104 cells.<br /> (Copyright © 2015 Elsevier B.V. All rights reserved.)
Details
- Language :
- English
- ISSN :
- 1879-0984
- Volume :
- 223
- Database :
- MEDLINE
- Journal :
- Journal of virological methods
- Publication Type :
- Academic Journal
- Accession number :
- 26235236
- Full Text :
- https://doi.org/10.1016/j.jviromet.2015.07.016