Development of a rapid agglutination latex test for diagnosis of enteropathogenic and enterohemorrhagic Escherichia coli infection in developing world: defining the biomarker, antibody and method.

Autor: Rocha LB; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil., Santos AR; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil., Munhoz DD; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil., Cardoso LT; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil., Luz DE; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil., Andrade FB; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil., Horton DS; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil., Elias WP; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil., Piazza RM; Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil.
Jazyk: angličtina
Zdroj: PLoS neglected tropical diseases [PLoS Negl Trop Dis] 2014 Sep 25; Vol. 8 (9), pp. e3150. Date of Electronic Publication: 2014 Sep 25 (Print Publication: 2014).
DOI: 10.1371/journal.pntd.0003150
Abstrakt: Background: Enteropathogenic and enterohemorrhagic Escherichia coli (EPEC/EHEC) are human intestinal pathogens responsible for diarrhea in both developing and industrialized countries. In research laboratories, EPEC and EHEC are defined on the basis of their pathogenic features; nevertheless, their identification in routine laboratories is expensive and laborious. Therefore, the aim of the present work was to develop a rapid and simple assay for EPEC/EHEC detection. Accordingly, the EPEC/EHEC-secreted proteins EspA and EspB were chosen as target antigens.
Methodology: First, we investigated the ideal conditions for EspA/EspB production/secretion by ELISA in a collection of EPEC/EHEC strains after cultivating bacterial isolates in Dulbecco's modified Eagle's medium (DMEM) or DMEM containing 1% tryptone or HEp-2 cells-preconditioned DMEM, employing either anti-EspA/anti-EspB polyclonal or monoclonal antibodies developed and characterized herein. Subsequently, a rapid agglutination latex test (RALT) was developed and tested with the same collection of bacterial isolates.
Principal Findings: EspB was defined as a biomarker and its corresponding monoclonal antibody as the tool for EPEC/EHEC diagnosis; the production of EspB was better in DMEM medium. RALT assay has the sensitivity and specificity required for high-impact diagnosis of neglected diseases in the developing world.
Conclusion: RALT assay described herein can be considered an alternative assay for diarrhea diagnosis in low-income countries since it achieved 97% sensitivity, 98% specificity and 97% efficiency.
Databáze: MEDLINE
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