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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2009.tde-29102009-083217
Document
Author
Full name
Lucas Miranda Marques
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2009
Supervisor
Committee
Timenetsky, Jorge (President)
Guimarães, Elizabeth Oliveira da Costa Freitas
Nascimento, Elmiro Rosendo do
Pellizari, Vivian Helena
Pinheiro, Ericka Tavares
Title in Portuguese
Estudo da variabilidade genética e dos fatores de virulência de isolados de Ureaplasma diversum.
Keywords in Portuguese
Ureaplasma diversum
Bactérias aeróbicas gram-negativas
Cápsulas
Fosfolipases C
Ig A protease
Imunoglobulinas
Invasão bacteriana
Microbiologia
Variação genética
Abstract in Portuguese
O presente trabalho teve como objetivo o estudo da variabilidade genética e dos fatores de virulência de isolados de U. diversum. As cepas foram submetidas a sequenciamento dos genes da urease e 16S rRNA e a testes para verificar os fatores de virulência: cápsula, fosfolipase C, IgAse e adesão e invasão. A análise do sequênciamento parcial do gene 16S rRNA resultou na presença de polimorfismos em 44 posições da seqüência, que diferenciou as amostras em sete grupos. Em relação aos fatores de virulência, os dados mostraram que as cepas estudadas apresentaram uma camada densa ao redor da membrana celular dos microrganismos e atividade de fosfolipase C. No entanto, não foi observado a atividade de IgAse nas cepas. Em relação a atividade de invasão, observou-se que os ureaplasma estudados puderam ser visualizados no interior de células Hep-2 com apenas um minutos de infecção, sendo observados em uma região perinuclear, mas não no interior do núcleo. Além disto, pode verificar que entre 1% a 10% dos ureaplasmas estudos penetraram na célula pelo teste da gentamicina.
Title in English
Study of genetic variability and virulence factors of Ureaplasma diversum isolates.
Keywords in English
Ureaplasma diversum
Aerobic gram-negative
Bacterial invasion
Capsules
Genetic variation
IgA protease
Immunoglobulins
Microbiology
Phospholipases C
Abstract in English
The aim of the present study was the study of genetic variability and virulence factors of U. diversum clinical isolates. The strains were submitted to sequencing for 16S rRNA and urease genes. Moreover, the strains were analyzed to the virulence factors: capsule, phospholipase C, IgA protease and adhesion and invasion into Hep-2 cells. The sequencing of parcial 16S rRNA gene showed polymorphic patterns into 44 positions. These polymorphisms clustered the strains in seven groups. For the virulence factors, ureaplasma cells showed a dense-stained external capsule-like structure surrounding the cell membrane. A high level of phospholipase C activity was also detected in 31 studied ureaplasma. However, no strains showed IgA protease activity. For the invasion assay, the isolates and strains used were detected inside the cells after infection of one minute. The invasions of the ureaplasmas surrounded the nuclear region but were not observed inside the nuclei. The gentamicin invasion assay detected that 1% to 10% of studied ureaplasmas were inside the infected cells.
 
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Publishing Date
2009-12-04
 
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