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

<h4>Background</h4>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; n...

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Autores principales:	Letícia B Rocha, Anna R R Santos, Danielle D Munhoz, Lucas T A Cardoso, Daniela E Luz, Fernanda B Andrade, Denise S P Q Horton, Waldir P Elias, Roxane M F Piazza
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270
Acceso en línea:	https://doaj.org/article/443550ba903f4a63a9845df6e30d701d
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spelling	oai:doaj.org-article:443550ba903f4a63a9845df6e30d701d2021-11-25T06:32:00ZDevelopment of a rapid agglutination latex test for diagnosis of enteropathogenic and enterohemorrhagic Escherichia coli infection in developing world: defining the biomarker, antibody and method.1935-27271935-273510.1371/journal.pntd.0003150https://doaj.org/article/443550ba903f4a63a9845df6e30d701d2014-09-01T00:00:00Zhttps://doi.org/10.1371/journal.pntd.0003150https://doaj.org/toc/1935-2727https://doaj.org/toc/1935-2735<h4>Background</h4>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.<h4>Methodology</h4>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.<h4>Principal findings</h4>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.<h4>Conclusion</h4>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.Letícia B RochaAnna R R SantosDanielle D MunhozLucas T A CardosoDaniela E LuzFernanda B AndradeDenise S P Q HortonWaldir P EliasRoxane M F PiazzaPublic Library of Science (PLoS)articleArctic medicine. Tropical medicineRC955-962Public aspects of medicineRA1-1270ENPLoS Neglected Tropical Diseases, Vol 8, Iss 9, p e3150 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270
spellingShingle	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Letícia B Rocha Anna R R Santos Danielle D Munhoz Lucas T A Cardoso Daniela E Luz Fernanda B Andrade Denise S P Q Horton Waldir P Elias Roxane M F Piazza 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.
description	<h4>Background</h4>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.<h4>Methodology</h4>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.<h4>Principal findings</h4>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.<h4>Conclusion</h4>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.
format	article
author	Letícia B Rocha Anna R R Santos Danielle D Munhoz Lucas T A Cardoso Daniela E Luz Fernanda B Andrade Denise S P Q Horton Waldir P Elias Roxane M F Piazza
author_facet	Letícia B Rocha Anna R R Santos Danielle D Munhoz Lucas T A Cardoso Daniela E Luz Fernanda B Andrade Denise S P Q Horton Waldir P Elias Roxane M F Piazza
author_sort	Letícia B Rocha
title	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.
title_short	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.
title_full	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.
title_fullStr	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.
title_full_unstemmed	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.
title_sort	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.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/443550ba903f4a63a9845df6e30d701d
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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.

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