| Full text | |
| Author(s): |
Rocha, Leticia B.
[1]
;
Santos, Anna R. R.
[1]
;
Munhoz, Danielle D.
[1]
;
Cardoso, Lucas T. A.
[1]
;
Luz, Daniela E.
[1]
;
Andrade, Fernanda B.
[1]
;
Horton, Denise S. P. Q.
[1]
;
Elias, Waldir P.
[1]
;
Piazza, Roxane M. F.
[1]
Total Authors: 9
|
| Affiliation: | [1] Inst Butantan, Lab Bacteriol, Sao Paulo - Brazil
Total Affiliations: 1
|
| Document type: | Journal article |
| Source: | PLoS Neglected Tropical Diseases; v. 8, n. 9 SEP 2014. |
| Web of Science Citations: | 7 |
| Abstract | |
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. (AU) | |
| FAPESP's process: | 12/02618-9 - Validation of an immunochromatographic assay for the diagnosis of diarrheagenic Escherichia coli |
| Grantee: | Anna Raquel Ribeiro dos Santos |
| Support Opportunities: | Scholarships in Brazil - Master |
| FAPESP's process: | 09/14845-7 - Immunodiagnosis of diarrheagenic Escherichia coli |
| Grantee: | Roxane Maria Fontes Piazza |
| Support Opportunities: | Regular Research Grants |