Development of a universal intimin antiserum and PCR primers

Miranda Batchelor, Stuart Knutton, Alfredo Caprioli, Veronika Huter, Mazlina Zanial, Gordon Dougan, Gad Frankel

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) constitute a significant risk to human health worldwide. A hallmark of both pathogens is their ability to produce characteristic attaching-and- effacing (A/E) lesions in intestinal epithelial cells. Genes encoding A/E lesion formation map to a chromosomal pathogenicity island termed the locus of enterocyte effacement (LEE). Intimin, an LEE-encoded bacterial adhesion molecule, mediates the intimate bacterium-host cell interaction characteristic of A/E lesions. On the basis of characterization of the C- terminal 280-amino-acid cell binding domain of intimin (Int280661- 939), four distinct Int280 types (types α, γ, and δ) have been identified. Importantly, Int280α and Int280β antisera specifically recognized their respective intimin types. Using a conserved region of the intimin molecule (Int388-667) and primers synthesized to generate the recombinant Int388-667, we have now generated universal intimin antiserum and PCR primers that are reactive with the different intimin types expressed by both human and animal A/E lesion-forming strains. Use of immunogold electron microscopy to visualize intimin on the surfaces of EPEC and EHEC strains revealed, in general, a uniform distribution on the bacterial cell surface. However, a filamentous staining pattern was observed with a few strains expressing intimin γ. Cloning of the intimin eae gene from one such strain (strain ICC57) into strain CVD206, an EPEC strain which harbors a null deletion in eae, produced a uniform intimin staining pattern indicating that, if the filamentous staining pattern defines a filamentous form of intimin γ, it is dependent upon the genetic background of the strain and is not a feature of the intimin molecule.

Original languageEnglish
Pages (from-to)3822-3827
Number of pages6
JournalJournal of Clinical Microbiology
Volume37
Issue number12
Publication statusPublished - 1 Dec 1999
Externally publishedYes

Cite this

Batchelor, M., Knutton, S., Caprioli, A., Huter, V., Zanial, M., Dougan, G., & Frankel, G. (1999). Development of a universal intimin antiserum and PCR primers. Journal of Clinical Microbiology, 37(12), 3822-3827.
Batchelor, Miranda ; Knutton, Stuart ; Caprioli, Alfredo ; Huter, Veronika ; Zanial, Mazlina ; Dougan, Gordon ; Frankel, Gad. / Development of a universal intimin antiserum and PCR primers. In: Journal of Clinical Microbiology. 1999 ; Vol. 37, No. 12. pp. 3822-3827.
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Batchelor, M, Knutton, S, Caprioli, A, Huter, V, Zanial, M, Dougan, G & Frankel, G 1999, 'Development of a universal intimin antiserum and PCR primers' Journal of Clinical Microbiology, vol. 37, no. 12, pp. 3822-3827.

Development of a universal intimin antiserum and PCR primers. / Batchelor, Miranda; Knutton, Stuart; Caprioli, Alfredo; Huter, Veronika; Zanial, Mazlina; Dougan, Gordon; Frankel, Gad.

In: Journal of Clinical Microbiology, Vol. 37, No. 12, 01.12.1999, p. 3822-3827.

Research output: Contribution to journalArticleResearchpeer-review

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Batchelor M, Knutton S, Caprioli A, Huter V, Zanial M, Dougan G et al. Development of a universal intimin antiserum and PCR primers. Journal of Clinical Microbiology. 1999 Dec 1;37(12):3822-3827.