Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFRGS |
Texto Completo: | http://hdl.handle.net/10183/180038 |
Resumo: | Enterotoxigenic Escherichia coli (ETEC) causes diarrhea in pigs, referred to as colibacillosis. The aim of this study was to optimize multiplex polymerase chain reaction (PCR) and immunohistochemistry (IHC) analyses of paraffin-embedded material to detect pathogenic E. coli strains causing colibacillosis in pigs. Multiplex PCR was optimized for fimbriae (F18, F4, F6, F5, and F41) and toxins (types A and B heat-stable toxins [STaP and STb], heat-labile toxin [LT], and type 2 Shiga toxin [STx2e]), and IHC was optimized for an anti-E. coli polyclonal antibody. Samples (132) from pigs received between 2006 and 2014 with clinical and histopathological diagnoses of colibacillosis were analyzed. E. coli was detected by IHC in 78.7%, and at least one virulence factor gene was detected in 71.2%. Pathogenic strains of ETEC with at least one fimbria and one toxin were detected in 40% of the samples in multiplex PCR. The most frequent virulence types were F18-STaP (7.5%), F18-STaP-STb (5.7%), and F4-STaP (3.8%). A statistically significant association was noted between virulence factors F4, F18, STaP, and STb and positive immunostaining results. Colibacillosis diagnosis through multiplex PCR and IHC of paraffin-embedded tissues is a practical approach, as samples can be fixed and stored for long periods before analysis. |
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Lorenzo, Cíntia deAndrade, Caroline Pinto deMachado, Verônica Silveira LuizBianchi, Matheus ViezzerRolim, Veronica MachadoCruz, Raquel Aparecida Sales daDriemeier, David2018-07-04T02:26:31Z20181229-845Xhttp://hdl.handle.net/10183/180038001069962Enterotoxigenic Escherichia coli (ETEC) causes diarrhea in pigs, referred to as colibacillosis. The aim of this study was to optimize multiplex polymerase chain reaction (PCR) and immunohistochemistry (IHC) analyses of paraffin-embedded material to detect pathogenic E. coli strains causing colibacillosis in pigs. Multiplex PCR was optimized for fimbriae (F18, F4, F6, F5, and F41) and toxins (types A and B heat-stable toxins [STaP and STb], heat-labile toxin [LT], and type 2 Shiga toxin [STx2e]), and IHC was optimized for an anti-E. coli polyclonal antibody. Samples (132) from pigs received between 2006 and 2014 with clinical and histopathological diagnoses of colibacillosis were analyzed. E. coli was detected by IHC in 78.7%, and at least one virulence factor gene was detected in 71.2%. Pathogenic strains of ETEC with at least one fimbria and one toxin were detected in 40% of the samples in multiplex PCR. The most frequent virulence types were F18-STaP (7.5%), F18-STaP-STb (5.7%), and F4-STaP (3.8%). A statistically significant association was noted between virulence factors F4, F18, STaP, and STb and positive immunostaining results. Colibacillosis diagnosis through multiplex PCR and IHC of paraffin-embedded tissues is a practical approach, as samples can be fixed and stored for long periods before analysis.application/pdfengJournal of veterinary science. Vol. 19, n. 1 (2018), p. 27-33Escherichia coli enterotoxigênicaColibacilose suinaFatores de virulênciaImuno-histoquímicaReação em cadeia da polimeraseEnteritisEnterotoxigenic Escherichia coliSwine diseasesVirulence factorsPiglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissuesEstrangeiroinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRGSinstname:Universidade Federal do Rio Grande do Sul (UFRGS)instacron:UFRGSORIGINAL001069962.pdf001069962.pdfTexto completo (inglês)application/pdf6313490http://www.lume.ufrgs.br/bitstream/10183/180038/1/001069962.pdfdaa3b3290bbba859357c224f06d95730MD51TEXT001069962.pdf.txt001069962.pdf.txtExtracted Texttext/plain35293http://www.lume.ufrgs.br/bitstream/10183/180038/2/001069962.pdf.txt3160008bddf16f54fa9158750c6bfcdaMD5210183/1800382018-07-05 02:25:14.518013oai:www.lume.ufrgs.br:10183/180038Repositório de PublicaçõesPUBhttps://lume.ufrgs.br/oai/requestopendoar:2018-07-05T05:25:14Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS)false |
dc.title.pt_BR.fl_str_mv |
Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues |
title |
Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues |
spellingShingle |
Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues Lorenzo, Cíntia de Escherichia coli enterotoxigênica Colibacilose suina Fatores de virulência Imuno-histoquímica Reação em cadeia da polimerase Enteritis Enterotoxigenic Escherichia coli Swine diseases Virulence factors |
title_short |
Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues |
title_full |
Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues |
title_fullStr |
Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues |
title_full_unstemmed |
Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues |
title_sort |
Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues |
author |
Lorenzo, Cíntia de |
author_facet |
Lorenzo, Cíntia de Andrade, Caroline Pinto de Machado, Verônica Silveira Luiz Bianchi, Matheus Viezzer Rolim, Veronica Machado Cruz, Raquel Aparecida Sales da Driemeier, David |
author_role |
author |
author2 |
Andrade, Caroline Pinto de Machado, Verônica Silveira Luiz Bianchi, Matheus Viezzer Rolim, Veronica Machado Cruz, Raquel Aparecida Sales da Driemeier, David |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Lorenzo, Cíntia de Andrade, Caroline Pinto de Machado, Verônica Silveira Luiz Bianchi, Matheus Viezzer Rolim, Veronica Machado Cruz, Raquel Aparecida Sales da Driemeier, David |
dc.subject.por.fl_str_mv |
Escherichia coli enterotoxigênica Colibacilose suina Fatores de virulência Imuno-histoquímica Reação em cadeia da polimerase |
topic |
Escherichia coli enterotoxigênica Colibacilose suina Fatores de virulência Imuno-histoquímica Reação em cadeia da polimerase Enteritis Enterotoxigenic Escherichia coli Swine diseases Virulence factors |
dc.subject.eng.fl_str_mv |
Enteritis Enterotoxigenic Escherichia coli Swine diseases Virulence factors |
description |
Enterotoxigenic Escherichia coli (ETEC) causes diarrhea in pigs, referred to as colibacillosis. The aim of this study was to optimize multiplex polymerase chain reaction (PCR) and immunohistochemistry (IHC) analyses of paraffin-embedded material to detect pathogenic E. coli strains causing colibacillosis in pigs. Multiplex PCR was optimized for fimbriae (F18, F4, F6, F5, and F41) and toxins (types A and B heat-stable toxins [STaP and STb], heat-labile toxin [LT], and type 2 Shiga toxin [STx2e]), and IHC was optimized for an anti-E. coli polyclonal antibody. Samples (132) from pigs received between 2006 and 2014 with clinical and histopathological diagnoses of colibacillosis were analyzed. E. coli was detected by IHC in 78.7%, and at least one virulence factor gene was detected in 71.2%. Pathogenic strains of ETEC with at least one fimbria and one toxin were detected in 40% of the samples in multiplex PCR. The most frequent virulence types were F18-STaP (7.5%), F18-STaP-STb (5.7%), and F4-STaP (3.8%). A statistically significant association was noted between virulence factors F4, F18, STaP, and STb and positive immunostaining results. Colibacillosis diagnosis through multiplex PCR and IHC of paraffin-embedded tissues is a practical approach, as samples can be fixed and stored for long periods before analysis. |
publishDate |
2018 |
dc.date.accessioned.fl_str_mv |
2018-07-04T02:26:31Z |
dc.date.issued.fl_str_mv |
2018 |
dc.type.driver.fl_str_mv |
Estrangeiro info:eu-repo/semantics/article |
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info:eu-repo/semantics/publishedVersion |
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publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10183/180038 |
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1229-845X |
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001069962 |
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http://hdl.handle.net/10183/180038 |
dc.language.iso.fl_str_mv |
eng |
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dc.relation.ispartof.pt_BR.fl_str_mv |
Journal of veterinary science. Vol. 19, n. 1 (2018), p. 27-33 |
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openAccess |
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