Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR

Detalhes bibliográficos
Autor(a) principal: Vinatea,Cecília E.B.
Data de Publicação: 2006
Outros Autores: Sincero,Thaís C.M., Simões,Claúdia M.O., Barardi,Célia R.M.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Microbiology
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822006000100012
Resumo: Shellfish are readily contaminated with viruses present in water containing sewage due to the concentrating effect of filter feeding. Enteroviruses are generally used as a model for the detection of viruses from shellfish due to their public health significance. In the present work, oysters were placed in glass aquaria containing seawater plus unicellular algae. Two experiments were performed: 1) oysters bioaccumulating four different poliovirus type 2 concentrations: 5 x 10(4), 2.5 x 10(4), 5 x 10³ and 5 x 10² PFU/mL during 20h; 2) oyster tissues directly inoculated with 6.0 x 10(5) and 1.0 x 10(5) PFU/mL. After viruses seeding, tissue samples were processed by an adsorption-elution-precipitation method. Positive controls were performed by seeding 6.0 x 10(5) PFU/mL of poliovirus type 2 directly on the final oyster tissue extracts. Oyster extracts were assayed for viruses recovery by plaque assay, RT-PCR and integrated cell culture-PCR methodologies (ICC/PCR). The last one was based on the inoculation of the samples onto VERO cell monolayer followed by RT-PCR analysis of the infected cell fluid. In the first experiment (20h bioaccumulation) until 5 x 10³ PFU were detected after 24 and 48h growth on VERO cells. Direct RT-PCR and ICC/PCR were able to detect 3 and 0.04 PFU of poliovirus, respectively, when bioaccumulation assay was used. When direct tissue virus seeding was performed, the plaque assays showed that polioviruses were recovered in all tested concentrations. Based on these results, it is possible to conclude that viable polioviruses can be detected in oysters after bioaccumulation and these techniques can be directly applied for monitoring virus contamination in environmental samples.
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spelling Detection of poliovirus type 2 in oysters by using cell culture and RT-PCRoysterspoliovirusbioaccumulationcell cultureRT-PCRShellfish are readily contaminated with viruses present in water containing sewage due to the concentrating effect of filter feeding. Enteroviruses are generally used as a model for the detection of viruses from shellfish due to their public health significance. In the present work, oysters were placed in glass aquaria containing seawater plus unicellular algae. Two experiments were performed: 1) oysters bioaccumulating four different poliovirus type 2 concentrations: 5 x 10(4), 2.5 x 10(4), 5 x 10³ and 5 x 10² PFU/mL during 20h; 2) oyster tissues directly inoculated with 6.0 x 10(5) and 1.0 x 10(5) PFU/mL. After viruses seeding, tissue samples were processed by an adsorption-elution-precipitation method. Positive controls were performed by seeding 6.0 x 10(5) PFU/mL of poliovirus type 2 directly on the final oyster tissue extracts. Oyster extracts were assayed for viruses recovery by plaque assay, RT-PCR and integrated cell culture-PCR methodologies (ICC/PCR). The last one was based on the inoculation of the samples onto VERO cell monolayer followed by RT-PCR analysis of the infected cell fluid. In the first experiment (20h bioaccumulation) until 5 x 10³ PFU were detected after 24 and 48h growth on VERO cells. Direct RT-PCR and ICC/PCR were able to detect 3 and 0.04 PFU of poliovirus, respectively, when bioaccumulation assay was used. When direct tissue virus seeding was performed, the plaque assays showed that polioviruses were recovered in all tested concentrations. Based on these results, it is possible to conclude that viable polioviruses can be detected in oysters after bioaccumulation and these techniques can be directly applied for monitoring virus contamination in environmental samples.Sociedade Brasileira de Microbiologia2006-03-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822006000100012Brazilian Journal of Microbiology v.37 n.1 2006reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S1517-83822006000100012info:eu-repo/semantics/openAccessVinatea,Cecília E.B.Sincero,Thaís C.M.Simões,Claúdia M.O.Barardi,Célia R.M.eng2006-05-19T00:00:00Zoai:scielo:S1517-83822006000100012Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2006-05-19T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false
dc.title.none.fl_str_mv Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR
title Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR
spellingShingle Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR
Vinatea,Cecília E.B.
oysters
poliovirus
bioaccumulation
cell culture
RT-PCR
title_short Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR
title_full Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR
title_fullStr Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR
title_full_unstemmed Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR
title_sort Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR
author Vinatea,Cecília E.B.
author_facet Vinatea,Cecília E.B.
Sincero,Thaís C.M.
Simões,Claúdia M.O.
Barardi,Célia R.M.
author_role author
author2 Sincero,Thaís C.M.
Simões,Claúdia M.O.
Barardi,Célia R.M.
author2_role author
author
author
dc.contributor.author.fl_str_mv Vinatea,Cecília E.B.
Sincero,Thaís C.M.
Simões,Claúdia M.O.
Barardi,Célia R.M.
dc.subject.por.fl_str_mv oysters
poliovirus
bioaccumulation
cell culture
RT-PCR
topic oysters
poliovirus
bioaccumulation
cell culture
RT-PCR
description Shellfish are readily contaminated with viruses present in water containing sewage due to the concentrating effect of filter feeding. Enteroviruses are generally used as a model for the detection of viruses from shellfish due to their public health significance. In the present work, oysters were placed in glass aquaria containing seawater plus unicellular algae. Two experiments were performed: 1) oysters bioaccumulating four different poliovirus type 2 concentrations: 5 x 10(4), 2.5 x 10(4), 5 x 10³ and 5 x 10² PFU/mL during 20h; 2) oyster tissues directly inoculated with 6.0 x 10(5) and 1.0 x 10(5) PFU/mL. After viruses seeding, tissue samples were processed by an adsorption-elution-precipitation method. Positive controls were performed by seeding 6.0 x 10(5) PFU/mL of poliovirus type 2 directly on the final oyster tissue extracts. Oyster extracts were assayed for viruses recovery by plaque assay, RT-PCR and integrated cell culture-PCR methodologies (ICC/PCR). The last one was based on the inoculation of the samples onto VERO cell monolayer followed by RT-PCR analysis of the infected cell fluid. In the first experiment (20h bioaccumulation) until 5 x 10³ PFU were detected after 24 and 48h growth on VERO cells. Direct RT-PCR and ICC/PCR were able to detect 3 and 0.04 PFU of poliovirus, respectively, when bioaccumulation assay was used. When direct tissue virus seeding was performed, the plaque assays showed that polioviruses were recovered in all tested concentrations. Based on these results, it is possible to conclude that viable polioviruses can be detected in oysters after bioaccumulation and these techniques can be directly applied for monitoring virus contamination in environmental samples.
publishDate 2006
dc.date.none.fl_str_mv 2006-03-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
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dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822006000100012
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822006000100012
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1517-83822006000100012
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
dc.source.none.fl_str_mv Brazilian Journal of Microbiology v.37 n.1 2006
reponame:Brazilian Journal of Microbiology
instname:Sociedade Brasileira de Microbiologia (SBM)
instacron:SBM
instname_str Sociedade Brasileira de Microbiologia (SBM)
instacron_str SBM
institution SBM
reponame_str Brazilian Journal of Microbiology
collection Brazilian Journal of Microbiology
repository.name.fl_str_mv Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)
repository.mail.fl_str_mv bjm@sbmicrobiologia.org.br||mbmartin@usp.br
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