Isolation, immunochemical demonstration of field strains of porcine group A rotaviruses and electrophoretic analysis of RNA segments of group A and C rotaviruses
2006
Smitalova, R.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) E-mail:[email protected] | Rodak, L.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) | Psikal, I.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) | Smid, B.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic))
The present study was focused on the isolation of rotaviruses from rotavirus electron positive samples of faeces on a susceptible tissue culture. We collected 195 faeces samples of diarrhoeic pigs and calves. Rotavirus was demonstrated by electron microscopy using the method of negative staining in 27 samples and by ELISA using monoclonal antibodies to the group antigen VP6 in 44 samples. Nine samples were selected for virus isolation. Three virus isolates (P375/4, P410/4 and P646/1) were successfully adapted to growth in cell line MA-104. These isolates were allocated to group A rotaviruses based on ELISA, immunoperoxidase test and electropherotype analysis. Electropherotype analysis demonstrated changes during passage in cell line in 2 of the 3 isolates. The selected sample P543/1 was negative in ELISA in a faecal sample. Electropherotype analysis of this sample revealed a "longer" electropherotype profile. The profile was suggestive of group C rotavirus. This was confirmed by RT-PCR and by sequence analysis.
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