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J Vet Sci ; 19(1): 27-33, 2018 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-28693311

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.


Assuntos
Diarreia/veterinária , Escherichia coli Enterotoxigênica/isolamento & purificação , Escherichia coli Enterotoxigênica/patogenicidade , Infecções por Escherichia coli/veterinária , Imuno-Histoquímica/veterinária , Reação em Cadeia da Polimerase Multiplex/veterinária , Doenças dos Suínos/diagnóstico , Animais , Diarreia/diagnóstico , Diarreia/microbiologia , Enterotoxinas/genética , Enterotoxinas/metabolismo , Infecções por Escherichia coli/diagnóstico , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/genética , Fímbrias Bacterianas/fisiologia , Imuno-Histoquímica/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Inclusão em Parafina/veterinária , Estudos Retrospectivos , Suínos , Doenças dos Suínos/microbiologia , Virulência , Fatores de Virulência/genética
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