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

Autor(es):Mostrar menos -

Menezes, M. A. ^[1] ; Rocha, L. B. ^[1] ; Koga, P. C. M. ^[1] ; Fernandes, I. ^[2] ; Nara, J. M. ^[1] ; Magalhaes, C. A. ^[1] ; Abe, C. M. ^[1] ; Ayala, C. O. ^[3] ; Burgos, Y. K. ^[3] ; Elias, W. P. ^[1] ; Castro, A. F. P. ^[3] ; Piazza, R. M. F. ^[1]

Afiliação do(s) autor(es)

^[1] Inst Butantan, Lab Bacteriol, BR-05503900 Sao Paulo - Brazil

^[2] Inst Butantan, Lab Imunopatol, BR-05503900 Sao Paulo - Brazil

^[3] Univ Sao Paulo, Inst Ciencias Biomed, Dept Microbiol, BR-05508 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Applied Microbiology; v. 108, n. 3, p. 878-887, MAR 2010.
• Citações Web of Science: 10

Resumo

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 center dot 3 x 10-8 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. (AU)

• Processo FAPESP: 04/12136-5 - Escherichia coli enteropatogênica atípica (EPEC atípica)
• Beneficiário: Waldir Pereira Elias Junior
• Linha de fomento: Auxílio à Pesquisa - Temático
• Processo FAPESP: 99/09458-0 - Desenvolvimento de testes imunosorológicos para detecção das toxinas ST e LT produzidas por Escherichia coli enterotoxigênica e das toxinas STX produzidas por Escherichia coli enterohemorrágica
• Beneficiário: Roxane Maria Fontes Piazza
• Linha de fomento: Auxílio à Pesquisa - Regular