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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Production and characterization of rabbit polyclonal sera against Shiga toxins Stx1 and Stx2 for detection of Shiga toxin-producing Escherichia coli

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Author(s):
Mendes-Ledesma, Meire R. B. [1] ; Rocha, Leticia B. [1] ; Bueris, Vanessa [1] ; Krause, Gladys [2] ; Beutin, Lothar [2] ; Franzolin, Marcia R. [1] ; Trabulsi, Luiz R. ; Elias, Waldir P. [1] ; Piazza, Roxane M. F. [1]
Total Authors: 9
Affiliation:
[1] Inst Butantan, Bacteriol Lab, BR-05503900 Sao Paulo - Brazil
[2] Fed Inst Risk Assessment, Natl Reference Lab Escherichia Coli, D-12277 Berlin - Germany
Total Affiliations: 2
Document type: Journal article
Source: MICROBIOLOGY AND IMMUNOLOGY; v. 52, n. 10, p. 484-491, OCT 2008.
Web of Science Citations: 12
Abstract

STEC has emerged as an important group of enteric pathogens worldwide. In this study, rabbit polyclonal Stx1 and Stx2 antisera were raised and employed in the standardization of immunoassays for STEC detection. Using their respective antisera, the limit of detection of the toxin was 35.0 pg for Stx1 and 5.4 pg for Stx2. By immunoblotting, these antisera recognized both toxin subunits. Cross-reactivity was observed in the A subunit, but only Stx2 antiserum was able to neutralize the cytotoxicity of both toxins in the Vero cell assay. Six stx-harboring E. coli isolates were analyzed for their virulence traits. They belonged to different serotypes, including the O48:H7, described for the first time in Brazil. Only three strains harbored eae, and the e-hly gene and hemolytic activity was detected in five strains. Three isolates showed new stx2 variants (stx(2v-ha) and stx(2vb-hb)). The ELISA assay detected all six isolates, including one VCA-negative isolate, while the immunodot assay failed to detect one isolate, which was VCA-positive. In contrast, the colony-immunoblot assay detected only one VCA-positive isolate. Our results demonstrate that among the immunoassays developed in this study, the immunodot, and particularly the ELISA, appear as perspective for STEC detection in developing countries. (AU)

FAPESP's process: 04/12136-5 - Atypical enteropathogenetic Escherichia coli (atypical EPEC)
Grantee:Waldir Pereira Elias Junior
Support Opportunities: Research Projects - Thematic Grants