Wild rabies virus detection by plaque assay from naturally infected brains in different species.
A simple, sensitive and specific plaque assay protocol for the detection of wild type rabies virus in different species is described using confluent monolayers of chicken embryo cells in 6-well plates. Plaques are produced after application of either agarose or Sephadex G-100 overlay onto cell monolayers and incubation for 96 h after virus infection at 37 degrees C. The parameters affecting plaque appearance include cell seeding concentration, overlay composition and time of incubation after infection. Optimal conditions are seeding at a concentration of 4 x 10(6) cell/cm(3), incubation at 37 degrees C in 5% CO(2) atmosphere during 96 h, using either 1% agarose or 2% Sephadex G-100 overlays. The described plaque assay would be a new valuable tool in conducting various quantitative investigations, since the chicken embryo cells are susceptible to rabies virus infection from all species studied.
Cardoso
Tereza
T
UNESP - Departamento de Apoio, Produção e Saude Animal, Curso de Medicina Veterinaria, Faculdade de Odontologia, Rua Clóvis Pestana, 793, 16.050-680 Araçatuba, São Paulo, Brazil. tcardoso@fmva.unesp.br
Pilz
Daniela
D
D000818:Animals; D000833:Animals, Suckling; D001681:Biological Assay / Q000662:veterinary; D001921:Brain / Q000821:virology*; D002415:Cats; D002417:Cattle; D002642:Chick Embryo; D002685:Chiroptera; D004285:Dogs; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D006736:Horses; D051379:Mice; D011818:Rabies / Q000175:diagnosis / Q000662:veterinary; D011820:Rabies virus / Q000254:growth & development* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013045:Species Specificity; D010948:Viral Plaque Assay / Q000379:methods / Q000662:veterinary*