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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2009.tde-02022010-100052
Document
Author
Full name
Claudia de Oliveira Ayala
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2009
Supervisor
Committee
Castro, Antonio Fernando Pestana de (President)
Carvalho, Vania Maria de
Franzolin, Marcia Regina
Piazza, Roxane Maria Fontes
Vaz, Tania Mara Ibelli
Title in Portuguese
Sorologia de antígenos flagelares de amostras de Escherichia coli Enteropatogênicas (EPEC) e E. coli produtoras da Toxina de Shiga (STEC) isoladas de diferentes animais e análise comparativa do gene fliC por PCR-RFLP.
Keywords in Portuguese
Escherichia coli
FliC
Flagelina
Microbiologia
Reação em cadeia por polimerase (PCR)
RFLP
Abstract in Portuguese
A espécie Escherichia coli constitui um grupo de bactérias tipicamente não patogênicas e que fazem parte do trato intestinal de humanos e animais. As amostras são sorotipadas com base em seus antígenos de superfície O (somático), H (flagelar) e K (capsular). O antígeno flagelar correspondente ao filamento é formado pela polimerização da flagelina, codificada pelo gene fliC. O presente trabalho empregou a técnica de PCR-RFLP para analisar os padrões de antígenos flagelares de 112 amostras de EPEC e STEC. Quatorze amostras não amplificaram o gene fliC, 17 tiveram seu antígeno flagelar determinado apenas por PCR-RFLP e 75 amostras tiveram seus antígenos flagelares confirmados por esta técnica. Três antígenos H com padrões irregulares foram clonados e sequenciados. Após o sequenciamento, inserções e remoções de nucleotídeos foram encontradas. Até o momento, poucos estudos utilizam um número abrangente de amostras de STEC e EPEC provenientes de diferentes animais para a determinação do antígeno H empregando a técnica de PCR-RFLP do gene fliC. De acordo com os resultados encontrados neste estudo, podemos concluir que a técnica de PCR-RFLP do gene fliC é mais rápida, menos trabalhosa e mais eficiente que a metodologia de sorotipagem clássica.
Title in English
Serology of flagellar antigens from strains of enteropathogenic Escherichia coli (EPEC) and Shiga Toxin producing E. coli (STEC) isolated from different animals and comparative analysis of the fliC gene by PCR-RFLP.
Keywords in English
Escherichia coli
FliC
Flagellin
Microbiology
Polymerase chain reaction (PCR)
RFLP
Abstract in English
The Escherichia coli species consists of a group of typically non-pathogenic bacteria present in the intestinal tract of humans and animals. Strains are serotyped according to their O (somatic), H (flagellar) and K (capsular) surface antigens, in order to distinguish these microorganisms from the non-pathogenic members of the intestinal microbiota. The flagellar antigen corresponding to the filament is formed by the polymerization of the flagellin, codified by the fliC gene. This study employed the PCR-RFLP technique to analyze flagellar antigen patterns from 112 EPEC and STEC strains. Fourteen strains have not amplified the fliC gene, 17 had their flagellar antigen determined only by the PCR-RFLP and 75 strains had their flagellar antigen confirmed by this technique. Three H antigens with irregular patterns were cloned and sequenced. After sequencing, insertions and deletions of nucleotides were discovered. So far, few studies used a significant number of STEC and EPEC strains originated from different animals to determine H antigens employing the PCR-RFLP technique of the fliC gene. According to the findings of this study, we assumed that PCR-RFLP of the fliC gene is faster, less laborious and more efficient than classic serotyping methodology.
 
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Publishing Date
2010-03-05
 
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