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Doctoral Thesis
DOI
https://doi.org/10.11606/T.6.2006.tde-12022007-104048
Document
Author
Full name
Gillian Alonso Arruda
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2006
Supervisor
Committee
Germano, Pedro Manuel Leal (President)
Damiani, Durval
Falcão, Mário Cícero
Matte, Maria Helena
Telles, Evelise Oliveira
Title in Portuguese
Perfil genotípico de cepas de Listeria monocytogenes isoladas de alimentos: análise crítica das técnicas de PCR e PFGE e importância para a Saúde Pública.
Keywords in Portuguese
16S
Internalina
Listeria monocytogenes
PCR
PFGE
Vigilância
Abstract in Portuguese
Devido à gravidade da manifestação clínica e pelas altas taxas de mortalidade em populações de risco, o controle e a prevenção da listeriose, doença causada pela L. monocytogenes, representa um importante desafio às autoridades sanitárias. A dificuldade de recuperar organismos envolvidos nos surtos, a sub-notificação e a demora na realização de análises convencionais são alguns dos fatores que retardam ou impedem intervenções em situações de surto. Com o advento da biologia molecular,novas técnicas têm sido empregadas para a identificação e tipificação da L. monocytogenes, com o intuito de contribuir com os estudos epidemiológicos e de vigilância sanitária de alimentos. O presente estudo visou confirmar a taxonomia e o perfil genotípico de 31 cepas isoladas de diversos tipos de carne, supostamente identificadas como L. monocytogenes, por intermédio das técnicas de Reação de Cadeia pela Polimerase (PCR), com o emprego de iniciadores de identificação dos genes 16S rRNA e Internalina A e B, para confirmação da espécie. Os resultados indicaram que apenas 20 (65%) das cepas foram confirmadas como sendo da espécie investigada. Foi realizada ainda a avaliação de similaridade nas 20 cepas confirmadas, através da Eletroforese em Campo Pulsado (PFGE), que revelou grande diversidade genotípica, caracterizada por 18 diferentes perfis, segundo o coeficiente de similaridade de Pearson. Os resultados demonstraram que as técnicas moleculares são de grande utilidade para a identificação e tipificação de L. monocytogenes, caracterizando-se como importantes ferramentas epidemiológicas.
Title in English
Genotype profile of cepas of would listeria monocytogenes isolated of foods: critical analysis of the techniques of PCR and PFGE and importance for the public health.
Keywords in English
Sanitary Surveillance
16S
Internalin
Listeria monocytogenes
PCR
PFGE
Abstract in English
Due to the seriousness of clinical manifestations and high rates of mortality of the listeriosis disease in populations at risk, control and prevention of this disease, caused by L. monocytogenes, represent an important challenge to sanitation authorities. Difficulties of recovering organisms involved in outbreaks; sub notification; and, the delay in accomplishing conventional analysis are some of the factors that slow-down or even prevent health interventions in outbreak occurrences. With the advent of the molecular biology, new techniques have been employed for identifying and typifying the L. monocytogenes aiming at contributing to studies on epidemiology and sanitary surveillance of foods. The present study aimed at ratifying the taxonomy and the genotypic profile of 31 isolated stocks of varied types of meats supposedly identified as L. monocytogenes by means of the Polymerase Chain Reaction (PCR) employing identification starters of genes 16 rRNA and A and B Internalin in order to confirm the species. Results indicated that only 20 (65.0%) of the stocks were confirmed as being from the investigated species. Another evaluation of similarity were also carried out along with the 20 stocks ratified through pulsed field gel electrophoresis (PFGE), which disclosed a great genotypic diversity characterized by 18 different profiles, according to the Pearson’s similarity coefficient. Results demonstrated that the molecular techniques are very helpful for identifying and typifying the L. monocytogenes, characterizing themselves as important epidemiological tools.
 
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tese_Gillian.pdf (1.04 Mbytes)
Publishing Date
2007-02-26
 
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