Identification of enteropathogenic and enterohaemorrhagic Escherichia coli strains by immunoserological detection of intimin.
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.
Menezes
M A
MA
 Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
Rocha
L B
LB
 Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
Koga
P C M
PCM
 Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
Fernandes
I
I
 Laboratório de Imunopatologia, Instituto Butantan, São Paulo, SP, Brazil.
Nara
J M
JM
 Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
Magalhães
C A
CA
 Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
Abe
C M
CM
 Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
Ayala
C O
CO
 Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
Burgos
Y K
YK
 Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
Elias
W P
WP
 Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
Castro
A F P
AFP
 Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
Piazza
R M F
RMF
 Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil.
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
D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal; D029968:Escherichia coli Proteins; D007106:Immune Sera; D007074:Immunoglobulin G; C073819:eaeA protein, E coli