title: Isolation and propagation of the animal rotaviruses in MA-104 cells--30 years of practical experience.
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.
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96A, 07743 Jena, Germany.
authors.forename: Peter H
authors.initials: PH
authors.lastname: Otto
authors.affiliation: Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Reetz
authors.affiliation: Ludwig-Maximilians-Universität München, Institut für Infektionsmedizin und Zoonosen, Infektions- und Seuchenmedizin, Veterinärstraße 13, 80539 München, Germany.
authors.forename: Werner
authors.initials: W
authors.lastname: Eichhorn
authors.affiliation: Justus-Liebig-Universität Gießen, Institut für Hygiene und Infektionskrankheiten der Tiere, Frankfurter Str. 87-89, 35392 Gießen, Germany.
authors.forename: Werner
authors.initials: W
authors.lastname: Herbst
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96A, 07743 Jena, Germany. Electronic address: mandy.elschner@fli.bund.de.
authors.forename: Mandy C
authors.initials: MC
authors.lastname: Elschner
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D003588:Cytopathogenic Effect, Viral; D005243:Feces / Q000821:virology*; D012401:Rotavirus / Q000254:growth & development* / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000662:veterinary* / Q000821:virology; D012692:Serial Passage; D019562:Viral Load; D014776:Virus Cultivation / Q000379:methods*
keywords: Cell culture; Diarrhoea; Isolation; MA-104 cells; Rotavirus; Trypsin