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Doctoral Thesis
DOI
https://doi.org/10.11606/T.17.2024.tde-11042024-095217
Document
Author
Full name
Carla de Oliveira Cardoso
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2023
Supervisor
Committee
Figueiredo, Luiz Tadeu Moraes (President)
Durigon, Edison Luiz
Lemos, Elba Regina Sampaio de
Santana, Rodrigo de Carvalho
Title in Portuguese
Diagnóstico etiológico em pacientes com infecção viral do sistema nervoso central, por PCR, no período de 2018 a 2022
Keywords in Portuguese
Adenovírus
Herpesvírus
Infecções virais
Meningoencefalites virais
SARS-Cov-2
Sistema nervoso central
Vírus encefalitogênicos
Abstract in Portuguese
Considerando que pouco se sabe sobre a etiologia de metade dos casos de meningoencefalite viral e tendo em vista a relevância clínica destas infecções, esta Tese visou a avaliar a etiologia de infecções virais no SNC, por reação em cadeia da polimerase (PCR) pesquisando por 28 patógenos, em 195 pacientes que frequentaram o Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto (HCFMRP/USP), de 01 de janeiro de 2018 a 31 de dezembro de 2022. Fez-se a extração de DNA e RNA das amostras de LCR e para diagnóstico de vírus de RNA, fez-se a transcrição reversa (RT) do RNA, para obter DNA complementar (DNAc). Os extratos de ácidos nucléicos e DNAc foram submetidos a: PCR em tempo real (qPCR) com kit comercial para detecção de patógenos comuns de SNC; PCR convencional desenvolvido in house para detecção de flavivírus e alfavírus; qPCR, utilizando kit comercial para SARS-CoV-2. Pesquisa de JCV também foi incluída no estudo. Os resultados dos testes virológicos foram correlacionados a resultados de celularidade e bioquímica liquórica. Também, dados clínico-epidemiológicos de todos os pacientes foram analisados. Dos 195 pacientes com suspeita de infecção viral do SNC, foram encontrados vírus no líquor de 65 pacientes (33,3% dos casos). Esta positividade viral mostrou-se mais elevada que a observada em outros estudos. Os vírus encontrados no líquor foram HAdV=14(20,5%), EBV=12(17,5%), HHV7=11(16,2%), SARS-CoV-2=9(13,2%), VZV=5(7,4%), HSV1=4(5,9%), CMV=4(5,9%), JCV=4(5,9%), HHV6=2(3%). HEV, EB19 e DENV3 tiveram 1(1,5%) paciente infectado. Observou-se maior ocorrência de infecções virais no SNC de pacientes mais jovens, com 0 a 18 anos (p<0,002647) e 19 a 35 anos (p<0,017061). Também, a imunodepressão foi observada como um fator de risco para as infecções virais do SNC (p<0,00785). Ainda, mostraram maior risco para infecção viral do SNC pacientes de atividade/ocupação no lar, em instituições, escolas, em privação de liberdade, ou aqueles que convivem sempre com as mesmas pessoas (p<0,0000593). Interessantemente, observou-se correlação do diagnóstico viral com líquor de baixa celularidade (p<0,00908). A positividade diagnóstica das infecções virais de SNC pareceu ser fortemente influenciada pela variedade de vírus pesquisados o que recomenda uma ampliação no número de patógenos que devem ser testados. Também, os resultados sugerem que possa estar havendo uma mudança no perfil etiológico das infecções virais do SNC em nosso meio.
Title in English
Etiologic diagnosis in patients with viral infection of central nervous system, by PCR, from 2018 to 2022
Keywords in English
Adenovirus
Central nervous system
Encephalitic viruses
Herpesvirus
SARS-CoV-2
Viral infections
Viral meningoencephalitis
Abstract in English
Considering that little is known about the etiology of half of the cases of viral meningoencephalitis and given the clinical relevance of these infections, this thesis aimed to evaluate the etiology of viral infections in the CNS through polymerase chain reaction (PCR) testing for 28 pathogens in 195 patients who attended the Ribeirão Preto Medical School Hospital (HCFMRP/USP) from January 1, 2018, to December 31, 2022. DNA and RNA were extracted from cerebrospinal fluid (CSF) samples, and for the diagnosis of RNA viruses, reverse transcription (RT) of RNA was performed to obtain complementary DNA (cDNA). Nucleic acid extracts and cDNA were subjected to the following tests: Real-time polymerase chain reaction (qPCR) using a commercial kit for the detection of common CNS pathogens; In-house developed conventional PCR for the detection of flaviviruses and alphaviruses; qPCR using a commercial kit for SARS-CoV-2 detection; Investigation of JC virus (JCV). The results of virological tests were correlated with cell counts and cerebrospinal fluid biochemistry. Additionally, clinical and epidemiological data of all patients were collected. Among the 195 patients with suspected viral CNS infections, viruses were found in the cerebrospinal fluid of 65 patients (33.3% of cases). This viral positivity was higher than that observed in other studies. The viruses found in the cerebrospinal fluid were: HAdV = 14(20.5%), EBV = 12(17.5%), HHV7 = 11(16.2%), SARS-CoV-2 = 9(13.2%), VZV = 5(7.4%), HSV1 = 4(5.9%), CMV = 4(5.9%), JCV = 4(5.9%), HHV6 = 2(3%), HEV, EB19, and DENV3 each had 1(1.5%) infected patient. A higher occurrence of viral CNS infections was observed in younger patients, aged 0 to 18 years (p<0.002647), and 19 to 35 years (p<0.017061). Immunodepression was also observed as a risk factor for viral CNS infections (p<0.00785). Furthermore, individuals engaged in home-based activities, institutions, schools, detention facilities, or those who consistently interacted with the same people were at a higher risk of CNS viral infections (p<0.0000593). Interestingly, a correlation was observed between viral diagnosis and cerebrospinal fluid with low cellularity (p<0.00908).
 
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Publishing Date
2024-07-11
 
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