title: Identification of enteropathogenic and enterohaemorrhagic Escherichia coli strains by immunoserological detection of intimin.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
  
  
  CONCLUSIONS
  The rabbit anti-intimin IgG-enriched fraction in immunoblotting analysis is a useful tool for EPEC and EHEC diagnoses.
authors.affiliation:  Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: M A
authors.initials: MA
authors.lastname: Menezes
authors.affiliation:  Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: L B
authors.initials: LB
authors.lastname: Rocha
authors.affiliation:  Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: P C M
authors.initials: PCM
authors.lastname: Koga
authors.affiliation:  Laboratório de Imunopatologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: I
authors.initials: I
authors.lastname: Fernandes
authors.affiliation:  Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: J M
authors.initials: JM
authors.lastname: Nara
authors.affiliation:  Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: C A
authors.initials: CA
authors.lastname: Magalhães
authors.affiliation:  Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: C M
authors.initials: CM
authors.lastname: Abe
authors.affiliation:  Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: C O
authors.initials: CO
authors.lastname: Ayala
authors.affiliation:  Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Y K
authors.initials: YK
authors.lastname: Burgos
authors.affiliation:  Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: W P
authors.initials: WP
authors.lastname: Elias
authors.affiliation:  Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: A F P
authors.initials: AFP
authors.lastname: Castro
authors.affiliation:  Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: R M F
authors.initials: RMF
authors.lastname: Piazza
mesh_terms: D018829:Adhesins, Bacterial / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D054324:Enterohemorrhagic Escherichia coli / Q000145:classification*; D054308:Enteropathogenic Escherichia coli / Q000145:classification*; D029968:Escherichia coli Proteins / Q000276:immunology*; D005260:Female; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting / Q000379:methods; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D051381:Rats; D012680:Sensitivity and Specificity
chemical_list: D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal; D029968:Escherichia coli Proteins; D007106:Immune Sera; D007074:Immunoglobulin G; C073819:eaeA protein, E coli