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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2012.tde-26112012-095546
Document
Author
Full name
Letícia Barboza Rocha
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Piazza, Roxane Maria Fontes (President)
Gaspari, Elizabeth Natal de
Guth, Beatriz Ernestina Cabilio
Oliveira, Maria Leonor Sarno de
Vaz, Tania Mara Ibelli
Title in Portuguese
Desenvolvimento e padronização de testes imunocromatográficos para diagnóstico de Escherichia coli produtora da toxina de Shiga (STEC) e Escherichia coli enterotoxigênica (ETEC).
Keywords in Portuguese
Escherichia coli
Anticorpos
Diarreia
Testes imunológicos
Toxinas
Abstract in Portuguese
Escherichia enterotoxigênica e Escherichia coli produtora da toxina de Shiga (STEC) são globalmente envolvidas em doenças diarreicas e produzem a toxina termolábil e a toxina de Shiga (Stx1 e Stx2), as quais são prejudiciais ao homem. O diagnóstico é uma importante ferramenta para o correto tratamento e controle de surtos. Assim, testes imunocromatográficos (ICs) para ETEC e STEC baseados na detecção das toxinas foram desenvolvidos e padronizados utilizando anticorpos policlonais conjugados ao ouro coloidal (PAbs) e seus correspondentes anticorpos monoclonais (MAbs) anti-LT (IgG2b, 4,9 x 10-11 M), anti-Stx1 (IgG1, 2,5 x 10-10 M) e anti-Stx2 (IgG1, 6,1 x 10-10 M). Os três testes ICs detectaram até 15,6 ng/mL de LT, 15,6 ng/mL de Stx1 e 62,5 ng/mL de Stx2 e não apresentaram reatividade cruzada com os isolados não produtores da respectiva toxina alvo. Estes resultados demonstram a viabilidade e aplicabilidade dos testes ICs como ferramentas no diagnóstico de E. coli diarreiogênicas.
Title in English
Development and standardization of immunochromatographic test for Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic Escherichia coli (ETEC) diagnosis.
Keywords in English
Escherichia coli
Antibodies
Diarrhea
Immunological tests
Toxins
Abstract in English
Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic Escherichia coli (ETEC) are worldwide involved in diarrheal diseases and produce the heat-labile (LT) and the Shiga (Stx1 and Stx2) toxins, which, cause injuries to man. The diagnosis is an important tool for the correct treatment and control of outbreaks. Thus, immunochromatographic assays (ICs) for ETEC and STEC based on toxins detection were developed and standardized using polyclonal antibodies-gold conjugated (PAbs) and their corresponding monoclonal antibodies (MAbs): anti-LT (IgG2b, 4.9 x 10-11 M), anti-Stx1 (IgG1, 2.5 x 10-10 M) and anti-Stx2 (IgG1, 6.1 x 10-10 M). The three ICs detect up to 15.6 ng/mL of LT, 15.6 ng/mL of Stx1 and 62.5 ng/mL of Stx2 and showed no cross-reaction with non-producers isolates of the respective target toxin. These results demonstrate the feasibility and applicability of the test ICs as diagnostic tools for diarrheagenic E. coli.
 
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Publishing Date
2012-12-18
 
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