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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2013.tde-19032014-160513
Document
Author
Full name
Rafaella Almeida Lima Nunes
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Domotor, Charlotte Marianna Hársi (President)
Martinez, Marina Baquerizo
Timenetsky, Maria do Carmo Sampaio Tavares
Title in Portuguese
Aplicação de técnicas moleculares no diagnóstico laboratorial complementar das infecções virais do sistema nervoso central no Hospital Universitário da USP.
Keywords in Portuguese
Adenovírus
Enterovírus
Meningite
Meningoencefalite
Reação em cadeia por polimerase
Sistema nervoso central
Abstract in Portuguese
Enterovírus (HEV), herpesvírus 1 e 2 (HHV-1 e HHV-2) e adenovírus (HAdV) são importantes agentes de infecções do SNC. Neste trabalho, técnicas moleculares foram aplicadas para a detecção destes vírus em quadros de infecção do SNC. Amostras de líquor foram colhidas de pacientes atendidos no HU-USP entre agosto e novembro/2010 e fevereiro/2012 a janeiro/2013. Através da Nested-PCR HEV foram detectados em 9,8% das amostras, HAdV em 2,5% e HHV-1 e 2 em 1,1%, além de 3 casos de coinfecção, 2 entre HEV e HHV, e 1 entre HEV e HAdV. O material genético viral foi extraído através dos métodos Qiaamp DNA Blood (Qiagen®) e MagMAXTM Viral RNA Isolation (Ambiom), e este último pareceu mais adequado à aplicação na rotina clínica. A análise quimiocitológica do líquor mostrou-se importante no direcionamento da conduta clínica, mas a detecção do vírus é fundamental para a conclusão do diagnóstico. A PCR em tempo real, cuja padronização foi iniciada neste trabalho, consiste em importante ferramenta para a utilização futura no diagnóstico complementar das infecções virais do SNC.
Title in English
Molecular techniques application for the complementary laboratory diagnosis of viral infections of the central nervous system, at the University Hospital of USP.
Keywords in English
Adenovirus
Central nervous system
Enterovirus
Meningitis
Meningoencephalitis
Polymerase chain reaction
Abstract in English
Enteroviruses (HEV), herpesviruses 1 and 2 (HHV-1 and HHV-2) and adenoviruses (HAdV) are important causative agents of infections of the CNS. In this study, molecular techniques were applied to the detection of these viruses. CSF samples were collected from patients treated at the University Hospital of USP, between August and November, 2010, and February 2012 and January 2013. By the Nested-PCR reaction, HEV were detected in 9.8% of the samples, HAdV in 2.5% and HHV-1 and 2 in 1.1%. There were 3 cases of coinfection: 2 with HEV and HHV and other with HEV and HAdV. The viral genetic materials were extracted by QIAamp DNA Blood kit (Qiagen®) and MagMAXTM Viral RNA Isolation (Ambiom), and the second one showed to be more suitable for the application in clinical diagnosis. The CSF chemocytologic analysis proved to be important in directing the clinical conduct, but the detection of viruses is essential for the diagnosis. The real time PCR, which standardization was initiated in this work, will be an important tool for complementary diagnosis of viral infections of the CNS.
 
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Publishing Date
2014-03-29
 
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