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Identification of enteropathogenic and enterohaemorrhagic Escherichia coli strains by immunoserological detection of intimin.
Menezes, M A; Rocha, L B; Koga, P C M; Fernandes, I; Nara, J M; Magalhães, C A; Abe, C M; Ayala, C O; Burgos, Y K; Elias, W P; Castro, A F P; Piazza, R M F.
Affiliation
  • Menezes MA; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
  • Rocha LB; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
  • Koga PCM; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
  • Fernandes I; Laboratório de Imunopatologia, Instituto Butantan, São Paulo, SP, Brazil.
  • Nara JM; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
  • Magalhães CA; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
  • Abe CM; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
  • Ayala CO; Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
  • Burgos YK; Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
  • Elias WP; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
  • Castro AFP; Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
  • Piazza RMF; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
J Appl Microbiol ; 108(3): 878-887, 2010 Mar.
Article in En | MEDLINE | ID: mdl-19709337
ABSTRACT

AIMS:

To evaluate the sensitivity and specificity of polyclonal and monoclonal antibodies (Mabs) against intimin in the detection of enteropathogenic and enterohaemorrhagic Escherichia coli isolates using immunoblotting. METHODS AND

RESULTS:

Polyclonal and Mabs against the intimin-conserved region were raised, and their reactivities were compared in enteropathogenic E. coli (EPEC) and enterohaemorrhagic E. coli (EHEC) isolates using immunoblotting analysis. In comparison with rat antiserum, rabbit anti-intimin IgG-enriched fraction had a stronger recognition pattern to a wide spectrum of intimin types in different EPEC and EHEC serotypes. On the other hand, murine monoclonal IgG2b specific to intimin, with dissociation constant of 1.3x10(-8) mol l(-1), failed in the detection of some of these isolates.

CONCLUSION:

All employed antibodies showed 100% specificity, not reacting with any of the eae-negative isolates. The sensitivity range was according to the employed antisera, and 97% for rabbit anti-intimin IgG-enriched fraction, followed by 92% and 78% sensitivity with rat antisera and Mab. SIGNIFICANCE AND IMPACT OF THE STUDY The rabbit anti-intimin IgG-enriched fraction in immunoblotting analysis is a useful tool for EPEC and EHEC diagnoses.
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Full text: 1 Collection: 01-internacional Health context: 3_ND Database: MEDLINE Main subject: Adhesins, Bacterial / Escherichia coli Proteins / Enteropathogenic Escherichia coli / Enterohemorrhagic Escherichia coli / Antibodies, Monoclonal Type of study: Diagnostic_studies / Evaluation_studies Limits: Animals Language: En Journal: J Appl Microbiol Year: 2010 Document type: Article
Fulltext
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Full text: 1 Collection: 01-internacional Health context: 3_ND Database: MEDLINE Main subject: Adhesins, Bacterial / Escherichia coli Proteins / Enteropathogenic Escherichia coli / Enterohemorrhagic Escherichia coli / Antibodies, Monoclonal Type of study: Diagnostic_studies / Evaluation_studies Limits: Animals Language: En Journal: J Appl Microbiol Year: 2010 Document type: Article