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Journal of Clinical Microbiology, March 1998, p. 662-668, Vol. 36, No. 3
Department of Biochemistry,
Received 10 July 1997/Returned for modification 15 October
1997/Accepted 19 November 1997
Intimins are outer membrane proteins expressed by enteric bacterial
pathogens capable of inducing intestinal attachment-and-effacement lesions. A eukaryotic cell-binding domain is located within a 280-amino-acid (Int280) carboxy terminus of intimin polypeptides. Polyclonal antiserum was raised against Int280 from enteropathogenic Escherichia coli (EPEC) serotypes O127:H6 and O114:H2
(anti-Int280-H6 and anti-Int280-H2, respectively), and Western blot
analysis was used to explore the immunological relationship between the
intimin polypeptides expressed by different clinical EPEC and
enterohemorrhagic E. coli (EHEC) isolates, a rabbit
diarrheagenic E. coli strain (RDEC-1), and
Citrobacter rodentium. Anti-Int280-H6 serum reacted strongly with some EPEC serotypes, whereas anti-Int280-H2 serum reacted
strongly with strains belonging to different EPEC and EHEC serotypes,
RDEC-1, and C. rodentium. These observations were confirmed
by using purified Int280 in an enzyme-linked immunosorbent assay and by
immunogold and immunofluorescence labelling of whole bacterial cells.
Some bacterial strains were recognized poorly by either antiserum
(e.g., EPEC O86:H34 and EHEC O157:H7). By using PCR primers designed on
the basis of the intimin-encoding eae gene sequences of
serotype O127:H6, O114:H2, and O86:H34 EPEC and serotype O157:H7 EHEC,
we could distinguish between different eae gene
derivatives. Accordingly, the different intimin types were designated
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Detection of Intimins
,
,
, and
, Four
Intimin Derivatives Expressed by Attaching and Effacing Microbial
Pathogens
,
,
, and
, respectively.
*
Corresponding author. Mailing address: Department of
Biochemistry, Imperial College of Science, Technology and Medicine,
Exhibition Rd., London SW7 2AZ, United Kingdom. Phone: 44-71-594-5253. Fax: 44-71-594-5255. E-mail: g.frankel{at}ic.ac.uk.
Journal of Clinical Microbiology, March 1998, p. 662-668, Vol. 36, No. 3
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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