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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2014.tde-08122014-085028
Document
Author
Full name
Eliane Pessoa da Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Dias, Waldely de Oliveira (President)
Gomez, José Gregorio Cabrera
Silva, Rosa Maria
Title in Portuguese
Desenvolvimento de teste rápido, usando "dipsticks", para diagnóstico de Streptococcus pneumoniae.
Keywords in Portuguese
Dipstick
Streptococcus pneumoniae
Diagnóstico
Partículas coloidais
Abstract in Portuguese
Doenças invasivas causadas por S. pneumoniae provocam cerca de 1,5 milhões de mortes anuais de crianças no mundo. Testes imunocromatográficos rápidos (dipsticks) são alternativa a métodos diagnósticos tradicionais, devido à fácil execução e baixo custo. Neste estudo foram desenvolvidos dipsticks para diagnóstico de S. pneumoniae utilizando: anticorpos de captura - monoclonais anti-pneumolisina recombinante (Ply) - AcMs D9-43 e E10-24, reativos contra 21 sorotipos prevalentes de S. pneumoniae; anticorpos de detecção (conjugados com diferentes partículas coloidais coloridas - corante têxtil, microesferas e nanopartículas de ouro) - Igs anti-Ply e anti-vacina pneumocócica celular (em desenvolvimento no Instituto Butantan) (anti-WCPV). Dipsticks com anti-Ply ou anti-WCPV conjugados com ouro ou microesferas coloidais detectaram a bactéria em todos os sorotipos prevalentes avaliados, entre 104 e 105 UFC/ml e pneumolisina nativa, em sobrenadantes de cultivo de S. pneumoniae, até uma concentração estimada 19,3 ng/ml e 9.7 ng/ml, respectivamente. Os resultados foram visualizados em aproximadamente 10 min.
Title in English
Development of rapid test using "dipsticks" for diagnosis of Streptococcus pneumoniae.
Keywords in English
Dipstick
Streptococcus pneumoniae
Colloidal particles
Diagnosis
Abstract in English
Invasive diseases caused by S. pneumoniae are responsible for about 1.5 million child deaths per year worldwide. Rapid Immunochromatographic tests (dipsticks) are alternative to traditional diagnostic methods due to easy execution and low cost. In this study, dipsticks have been developed for diagnosis of S. pneumoniae using: capture antibodies - monoclonal anti- recombinant pneumolysin (Ply), MAbs D9 -43 and E10 - 24, reactive against 21 prevalent serotypes of S. pneumoniae; detection antibodies (conjugated with different colored colloidal particles - textile dye; microspheres and gold nanoparticles) - anti-Ply and anti whole cell pneumococcal vaccine (in developing in the Butantan Institute) (anti- WCPV). Dipsticks with anti-Ply or anti-WCPV conjugated with colloidal gold or microspheres detected the bacterium in all the evaluated prevalent serotypes, between 104 e 105 CFU/ml, and native pneumolysin in culture supernatants of S. pneumoniae to an estimated concentration 19.3 ng/ml and 9.7 ng/ml, respectively. The results could be visualized in around 10 min.
 
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Publishing Date
2014-12-10
 
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