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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Perfil de sensibilidade e genotipagem de leveduras isoladas de pacientes com candidemia em dois Hospitais de ReferÃncia TerciÃria de Fortaleza-Cearà / Sensitivety profile to antifulgal and evaluate the genotypical profile of the yeast isolated from candidemic patients tertiary care hospital from Northeast Brazil

DÃlia JÃssica Astete Medrano 24 September 2004 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / As infecÃÃes fÃngicas sistÃmicas causadas por leveduras do gÃnero Candida sÃo consideradas micoses oportunistas de alto risco em ambientes hospitalares. Essas infecÃÃes representam importante desafio terapÃutico, em razÃo do surgimento de espÃcies resistentes a antifÃngicos, associados a altos Ãndices de mortalidade. Por esta razÃo, este trabalho objetivou: Verificar a freqÃÃncia de fungemias no Hospital Geral Dr. CÃsar Cals (HGCC) e Hospital Infantil Albert Sabin (HIAS); identificar os agentes etiolÃgicos implicados; determinar o perfil de sensibilidade aos antifÃngicos e avaliar o perfil genotÃpico das espÃcies isoladas de pacientes com quadros de recorrÃncia. Para isso foram recuperadas, de 4 627 hemoculturas, 55 hemoculturas positivas para leveduras do HGCC, e, de 5 316 hemoculturas, 87 positivas para leveduras do HIAS. Destas, 23 do HGCC e 43 pacientes do HIAS que possuÃam histÃrias clÃnicas com dados completos entraram neste estudo. O diagnÃstico micolÃgico foi realizado por meio das caracterÃsticas bioquÃmicas e morfolÃgicas dos agentes etiolÃgicos isolados. O teste de sensibilidade aos antifÃngicos - anfotericina B, fluconazol, itraconazol e cetoconazol - utilizou o mÃtodo de microdiluiÃÃo em caldo, descrito no documento M27-A2 do NCCLS. As tÃcnicas de genotipagem foram realizadas por eletroforese em campo pulsÃtil (PFGE) e amplificaÃÃo aleatÃria do DNA (RAPD). Entre o perÃodo do junho do 2000 a junho do 2002, no HGCC, foram analisadas 4 627 hemoculturas, sendo positivas 1051, das quais 55 (5,2%) foram positivas para leveduras do gÃnero Candida. Em contrapartida, no perÃodo de junho do 2001 a junho do 2002, no HIAS, foram analisadas 5316 hemoculturas, sendo positivas 1520 amostras, das quais 87 (5,72%) foram positivas para leveduras dos gÃneros Candida e Rhodotorula. A principal espÃcie envolvida, dentro do gÃnero Candida, foi a C. parapsilosis com 36% e 42% no HGCC e HIAS, respectivamente. Os principais fatores de risco foram: a antibioticoterapia prÃvia (n=19; 90%), cateter venoso central, nutriÃÃo parenteral, sondagem gÃstrica , ventilaÃÃo mecÃnica, cirurgia e prematuridade. Quanto à resistÃncia a drogas antifÃngicas, foi observado que anfotericina B apresentou um Ãndice de resistÃncia de 4%, fluconazol- 52%, itraconazol- 50% e cetoconazol- 86%. A tÃcnica do PFGE caracterizou um perfil cromossÃmico de 6 bandas para C. parapsilosis e 4 para C. tropicalis. Esta tÃcnica permitiu a diferenciaÃÃo de uma cepa com padrÃo de bandas alterado para o Ãltimo episÃdio do paciente 4. A tÃcnica de RAPD caracterizou um padrÃo de distribuiÃÃo de bandas predominante para C. parapsilosis e foi capaz de caracterizar 4 perfis genÃmicos, indicando re-infecÃÃo em 3 pacientes e 2 perfis genÃmicos foram encontrados para C. tropicalis . Este ensaio demonstra a emergÃncia das Candida nÃo-albicans, especialmente a C. parapsilosis, como principal responsÃvel pelos casos de candidemias em dois hospitais de indicaÃÃo terciÃria de Fortaleza, associado a uma alta resistÃncia in vitro aos antifÃngicos. A variabilidade genÃtica encontrada nas C. parapsilosis e C. tropicalis, mediante as tÃcnicas do PFGE e RAPD, revelaram quadros de re-infecÃÃo, os quais erguem a possibilidade de que estas sejam decorrentes de uma contaminaÃÃo entre pacientes ou indiretamente, por intermÃdio de trabalhadores da saÃde. / The systemic infections caused by yeasts of the genus Candida are considered opportunist mycoses of high risk in hospital environments. These infections represent important therapeutical challenge, by reason of arising of species resistent to some antifungals, associated with high rate of mortality. Therefore, this work sought: To verify the frequency of fungemias at Dr. CÃsar Cals Hospital and Albert Sabin Children Hospital; identify the etiological agents implicated; determine the sensitivety profile to antifulgal and evaluate the genotypical profile of the species isolated from patients with recurrency. For this, were recuperated from 4627 hemcultures, 55 yeast-positive hemocultures at HGCC and from 5316 hemocultures 87 yeast-positive hemocultures at HIAS. From them, 23 patients at HGCC and from 43 at HIAS that had clinical histories with complet data entered this study. The mycological diagnosis was performed through biochemical and morphological characteristics of the etiological isolated. The test of sensitivety to antifungal- amphotericin B, fluconazole, itraconazole and cetoconazole- utilized the microdilution method in broth, reported in the document M27-A2 of the NCCLS. The genotyping techniques were performed through PFGE (Pulsed Field Gel Electrophresis), RAPD (Randomly amplified polymorphic DNA analysis). Within the period from june of 2000, to june of 2002 at HGCC, were analysed 4627 hemocultures, being positives 1051, of which 55 (5,2%) were positive for yeast of the genus Candida. On the other hand, in the period from june of 2001 to july of 2002 at HIAS, were analysed 5316 hemocultures, of which 87 (5,72%) were yeast-positive of the genus Candida and Rhodotorula.The main specie involved in the genus Candida, was C. parapsilosis with 36% and 42% of the cases at HGCC and HIAS respectively. The main risk factors associated with candidemia, were: the previous antibiotic therapy, central venous catheter, parenteral nutrition, gastric probe, mechanical ventilation, surgery and prematurity. About the resistance to antifungals drugs was observed that amphotericin B showed a resistance level of 4%, fluconazole-56%, itraconazole-52% and cetoconazole- 86%. The PFGE technique caracterized a cromossomic profile of 6 bands for C. parapsilosis and 4 for C. tropicalis. This technique permited the diferentiation of one cepa with changed pattern of bands for the last episode of the patient 4. The technique of RAPD chacacterized a distribution pattern of predominant bands for C. parapsilosis and was capable of characterize 4 genetical profiles, denoting reinfection in 3 patients and 2 genomic profiles were found for C. tropicalis. This assay shows the emergency of the non- albicans Candida, specially the C. parapsilosis, as main responsible for the cases of candidemia in the two hospitals of tertiary indication of Fortaleza, associated with a high resistance in vitro to antifungals. The genetical variability found in C. parapsilosis and C. tropicalis through the PFGE and RAPD techniques, revealed reinfection pictures, which rises the possibility that these be due to a contamination among patients or indirectly through the healthy workers.

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