Simian rotavirus SA-11 plaque formation in the presence of trypsin.
Incorporation of 5 micrograms of trpsin per ml of the overlay (Eagle minimal essential medium-0.7% Ionagar no. 2) was found to be necessary for plaque formation by simian rotavirus SA-11. Plaques of 3 to 4 mm in diameter were produced in MA-104 cells after 5 days of incubation at 37 degrees C. Plaque size was even larger (5 to 6 mm) in monolayers of African green monkey kidney cells. Addition of diethyl-aminotheyl-dextran, protamine sulfate, or 5-bromodeoxyuridine to the trypsin-containing overlay did not improve plaque formation by the virus. Incorporation of beef extract or yeast extract to a final concentration of 0.5% in the trypsin-containing overlay inhibited plaque formation. On the other hand, the presence of lactalbumin hydrolysate or peptone at a similar concentration in the overlay did not inhibit plaque formation. When methylcellulose was used instead of the agar as the solidifying agent in the overlay, no plaques were seen. SA-11 is a useful model for the study of human rotaviruses, and this relatively simple plaque assay system should further enhance its usefulness in this regard.
D000818:Animals; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002460:Cell Line; D003470:Culture Media; D003588:Cytopathogenic Effect, Viral; D003911:Dextrans / Q000494:pharmacology; D000882:Haplorhini; D007668:Kidney; D008747:Methylcellulose / Q000494:pharmacology; D011479:Protamines / Q000494:pharmacology; D012328:RNA Viruses / Q000187:drug effects*; D012401:Rotavirus / Q000187:drug effects* / Q000254:growth & development; D013696:Temperature; D014357:Trypsin / Q000494:pharmacology*; D014779:Virus Replication
D003470:Culture Media; D003911:Dextrans; D011479:Protamines; D008747:Methylcellulose; D014357:Trypsin; D001973:Bromodeoxyuridine