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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2009.tde-02122009-110053
Document
Author
Full name
Daniela Carvalho dos Santos
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2009
Supervisor
Committee
Harsi, Charlotte Marianna (President)
Gilio, Alfredo Elias
Pinho, João Renato Rebello
Silva, Edson Elias da
Zanotto, Paolo Marinho de Andrade
Title in Portuguese
Estabelecimento de métodos moleculares para aplicação no diagnóstico rápido de virus neurotrópicos.
Keywords in Portuguese
Adenovírus
Biologia molecular
Enterovírus
Herpesvírus 3 humano
Meningite viral (diagnóstico)
Microbiologia
Abstract in Portuguese
Diversos agentes virais são causadores de meningites e meningoencefalites. Neste estudo, técnicas moleculares foram utilizadas para detecção de HEV, HHV e HAdV em amostras de líquor colhidas de janeiro de 2005 a março de 2007. Dos três métodos de extração de DNA e RNA testados, o kit DNA Qiablood Qiagen® se mostrou o mais sensível e específico. A nested PCR detectou HEV em 28% das amostras, HSV em 4%, HHV-3 em 1%; HHV-4 em 0,3%, HHV-5 em 0,3%, HHV-6 em 0,7% e HAdV em 13%. Através da PCR em tempo real os HEV foram detectados em 23,3% e HSV em 5,1%. Por neutralização, somente duas amostras foram sorotipadas (Echovirus 6 e Coxsackievirus B). Os HEV detectados foram então seqüenciados para a determinação do sorotipo. Os sorotipos Echovirus 18 (53%) e Coxsackievirus B5 (26%) foram os mais freqüentes. As técnicas de biologia molecular aplicadas na detecção de HEV, HHV e HAdV no líquor trazem grandes vantagens ao diagnóstico de doenças do SNC graças à rapidez no diagnóstico, alta sensibilidade e especificidade.
Title in English
The integration of molecular methods into the rapid laboratorial diagnostic of neurotropic viruses.
Keywords in English
Adenovirus
Enterovirus
Herpesvirus 3 human
Microbiology
Molecular biology
Viral meningitis (diagnosis)
Abstract in English
Several viruses are etiological agents of meningitis and meningoencephalitis. In this study, molecular techniques were used for the detection of HEV, HHV and HAdV in liquor samples, collected from January 2005 to March 2007. Among the three methods tested for the extraction of DNA and RNA, the DNA Qiablood kit - Qiagen® was the most sensible and specific. HEV (28%), HSV (4%), HHV-3 (1%), HHV-4 (0.3%), HHV-5 (0.3%), HHV-6 (0.7%) and HAdV (13%) were detected by Nested PCR in the samples. By real time PCR, HEV were detected in 23.3% and HSV in 5.1%. Only two HEV could be serotyped by neutralization (Echovirus 6 and Coxsackievirus B). All detected HEV were then sequenced to determine the serotype. The serotypes echovirus 18 (53%) and Coxsackievirus B5 (26%) were the most frequent. Molecular biology techniques applied in the detection of HEV, HAdV and HHV in CSF bring major benefits to the diagnosis of meningitis thanks to the rapid diagnosis, high sensibility and specificity.
 
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Publishing Date
2010-03-02
 
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