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(es)

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]

Afiliação do(s) autor(es)

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

• Tipo de documento: Artigo Científico
• Fonte: PLoS Neglected Tropical Diseases; v. 8, n. 9 SEP 2014.
• Citações Web of Science: 7

Resumo

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)

• Processo FAPESP: 12/02618-9 - Validação de um ensaio imunocromatográfico para o diagnóstico de Escherichia coli diarreiogênica
• Beneficiário: Anna Raquel Ribeiro dos Santos
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 09/14845-7 - Imunodiagnóstico de Escherichia coli diarreiogênica
• Beneficiário: Roxane Maria Fontes Piazza
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 07/58610-8 - Escherichia coli diarreiogênica: ferramentas inovadoras para imunodiagnóstico
• Beneficiário: Letícia Barboza Rocha
• Modalidade de apoio: Bolsas no Brasil - Doutorado