Isolation of bovine coronavirus (BCoV) in monolayers of HmLu-1 cells
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Data
2005-09-01
Autores
Jerez, José Antonio
Gregori, Fábio
Brandão, Paulo Eduardo
Rodriguez, Cesar Alejandro Rosales
Ito, Fumio Honma
Buzinaro, Maria da Glória [UNESP]
Sakai, Takeo
Título da Revista
ISSN da Revista
Título de Volume
Editor
Sociedade Brasileira de Microbiologia
Resumo
A partir de 20 amostras fecais de bezerros, com quadro clínico de diarréia, positivas para BCoV por hemaglutinação/inibição da hemaglutinação (HA/HI), procedeu-se o isolamento viral em monocamadas de células da linhagem HmLu-1. Até a quinta passagem seriada, 13 apresentaram efeito citopático do tipo sincicial, semelhante à amostra padrão Kakegawa de BCoV. Ao serem submetidas a uma reação de soroneutralização com gamaglobulina anti-coronavírus bovino, 8 delas foram consideradas positivas, uma vez que o efeito citopático foi neutralizado. Ao serem tituladas e submetidas à reação de soroneutralização em microplacas, apenas 3 delas puderam ser confirmadas como positivas. As células da linhagem HmLu-1 mostraram-se permissivas para o isolamento de BCoV, todavia a baixa intensidade na replicação viral demonstrou ser necessário o desenvolvimento de novas metodologias para se poder alcançar esse intuito e a confirmação do isolamento por soroneutralização.
Isolation of BCoV was performed in monolayers of HmLu-1 cells, using 20 fecal samples from clinical cases of diarrhea in calves. Samples were positive for BCoV by means of hemagglutination / hemagglutination inhibition (HA/HI). Up to the fifth serial passage, 13 of these isolates presented syncytial cytopathic effect, similar to Kakegawa standard strain. When isolates were submitted to seroneutralization with gammaglobulin anti-bovine coronavirus, 8 of them were considered to be positive, once cytopathic effect was neutralized. After titration and seroneutralization in microplates, only three of them were confirmed as positive. The lineage HmLu-1 showed higher permissivity to BCoV isolation, but the low intensity of viral replication demonstrated that new methodologies should be developed for this purpose and then submitted to confirmation of isolation by means of seroneutralization.
Isolation of BCoV was performed in monolayers of HmLu-1 cells, using 20 fecal samples from clinical cases of diarrhea in calves. Samples were positive for BCoV by means of hemagglutination / hemagglutination inhibition (HA/HI). Up to the fifth serial passage, 13 of these isolates presented syncytial cytopathic effect, similar to Kakegawa standard strain. When isolates were submitted to seroneutralization with gammaglobulin anti-bovine coronavirus, 8 of them were considered to be positive, once cytopathic effect was neutralized. After titration and seroneutralization in microplates, only three of them were confirmed as positive. The lineage HmLu-1 showed higher permissivity to BCoV isolation, but the low intensity of viral replication demonstrated that new methodologies should be developed for this purpose and then submitted to confirmation of isolation by means of seroneutralization.
Descrição
Palavras-chave
bovine coronavirus (BCoV), Isolation, HmLu-1 cells, coronavírus bovino (BCoV), Isolamento, células HmLu-1
Como citar
Brazilian Journal of Microbiology. Sociedade Brasileira de Microbiologia, v. 36, n. 3, p. 207-210, 2005.