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| Author(s): |
Letícia Barboza Rocha
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) |
| Defense date: | 2008-04-15 |
| Examining board members: |
Roxane Maria Fontes Piazza;
Waldir Pereira Elias Junior;
Beatriz Ernestina Cabilio Guth
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| Advisor: | Roxane Maria Fontes Piazza |
| Abstract | |
STEC infection is a public health problem, causing hemolytic-uremic syndrome and haemorrhagic colitis. The detection of STEC isolates depends on a sensitive, specific and low cost diagnostic method. Thus, the aim of this study was to develop a capture ELISA using polyclonal and monoclonal antibodies. For this purpose, first of all a growth media that induce better expression and production of Stx was defined and monoclonals anti-Stx1 e anti-Stx2 antibodies were produced. The results show that the growth of the isolates for 4hs in EC broth with ciprofloxacin increased the production of Stx in the bacterial supernantant, and the monoclonal anti-Stx1antibody produced was able to recognize the A subunit of both toxins, besides showing higher affinity constant than anti-Stx2. Thus, the capture ELISA was standardized using 250 mg/ml of the IgG enriched fraction of rabbit anti-Stx1 sera and anti-Stx2 for coating and 2,5 mg/ml of monoclonal anti-Stx1 antibody for the capture of the toxin, showing 80,7% of sensibility and e 100% de specificity. (AU) | |