Identification of enteropathogenic and enterohaemorrhagic Escherichia coli strains by immunoserological detection of intimin

J Appl Microbiol. 2010 Mar;108(3):878-87. doi: 10.1111/j.1365-2672.2009.04484.x. Epub 2009 Aug 20.

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.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesins, Bacterial / immunology*
  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity
  • Enterohemorrhagic Escherichia coli / classification*
  • Enteropathogenic Escherichia coli / classification*
  • Escherichia coli Proteins / immunology*
  • Female
  • Immune Sera / immunology
  • Immunoblotting / methods
  • Immunoglobulin G / immunology
  • Mice
  • Mice, Inbred BALB C
  • Rabbits
  • Rats
  • Sensitivity and Specificity

Substances

  • Adhesins, Bacterial
  • Antibodies, Monoclonal
  • Escherichia coli Proteins
  • Immune Sera
  • Immunoglobulin G
  • eaeA protein, E coli