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Rev. chil. infectol ; 35(5): 601-605, 2018. tab
Article in Spanish | LILACS | ID: biblio-978076


Resumen La detección de virus en el líquido cefalorraquídeo (LCR) en pacientes infectados por VIH con carga viral (CV) indetectable en el plasma se ha denominado escape viral. Estas fugas pueden ser asintomáticas o asociadas con enfermedad neurológica. La discordancia de la carga viral de VIH entre plasma y LCR evidenciaría la presencia de distintos compartimentos del virus, con la posibilidad de identificar quasiespecies con mutaciones específicas que confieran resistencia a la TARV. Presentamos el caso clínico de un paciente con infección por VIH en etapa SIDA y una tuberculosis diseminada que presentó un cuadro neurológico manifestado por cefalea y un síndrome convulsivo, en que se encontró una discordancia entre la CV para VIH en plasma y LCR. El estudio genotípico del virus obtenido del LCR identificó nuevas mutaciones que determinaron un cambio de la TARV, con evolución posterior satisfactoria.

Detection of virus in cerebrospinal fluid (CSF) in HIV-infected patients with HIV viral load (VL) undetectable in plasma has been termed viral escape. These leaks may be asymptomatic from a neurological point of view, similar to plasma blips, or associated with neurological disease, with discordant VL between plasma and CSF, and may be evidence of a compartmentalization of the virus and the possibility of identifying quasispecies with mutations that confer resistance to ART. We present the case of a man with AIDS and disseminated tuberculosis who presented neurological symptomatology evidenced by headache and convulsive syndrome, who presented a discordance between plasma and CSF HIV VL; the genotypic test of the virus, obtained by lumbar puncture, identified new mutations that determined a change in ART with subsequent satisfactory evolution.

Humans , Male , Adult , Tuberculosis, Meningeal/diagnosis , HIV Infections/cerebrospinal fluid , Cerebrospinal Fluid/virology , HIV-1/genetics , Viral Load , Tuberculosis, Meningeal/complications , RNA, Viral/cerebrospinal fluid , HIV Infections/complications , Mutation/genetics
Arq. neuropsiquiatr ; 74(10): 810-815, Oct. 2016. tab, graf
Article in English | LILACS | ID: lil-796838


ABSTRACT The presence of hemoglobin in samples are considered an important inhibitory factor for polymerase chain reaction (PCR). The aim of this study was to examine the influence of red blood cells (RBC)s in cerebrospinal fluid (CSF) as an inhibitory factor to reverse transcription polymerase chain reaction (RT-PCR) for enteroviruses (EV). Forty-four CSF samples from patients showing characteristics of viral meningitis were assessed for EV by RT-PCR. Viral RNA extracted with guanidine isothyocianate buffer and virus detection was performed by in-house nested PCR. Positivity for EV RT-PCR was higher in CSF samples without RBCs than in samples with RBCs: 13(26%) and 36(9.2%), p = 0.001. In the group with positive EV RT-PCR, the mean + SD CSF RBC was 37 ± 183 cell/mm3; the group with negative results had 580 + 2,890 cell/mm3 (p = 0.007). The acceptable upper limit for CSF RBCs that could not influence RT-PCR was 108 cells/mm3. CSF samples with negative results for EV RT-PCR have more erythrocytes.

RESUMO A presença de hemoglobina em amostras de fluidos corporais é considerada um fator inibitório importante da reação em cadeia da polimerase (PCR). O objetivo deste estudo era examinar a influencia de hemácias no líquido cefalorraquidiano (LCR) como um fator inibitório da RT-PCR para enterovirus (EV). Quatrocentos e quarenta amostras de LCR de pacientes com características de meningite viral foram avaliados para enterovirus por RT-PCR. RNA viral foi extraído com tampão de isotiocianato de guanidina e a detecção viral foi feita com nested PCR in-house. A positividade do EV RT-PCR no LCR foi maior nas amostras de LCR sem hemácias do que as amostras com hemácias: 13 (26%) e 36 (9,2%), respectivamente (p = 0,001). No grupo com resultados EV RT-PCR positivo, a media ± DP do número de hemácias no LCR foi 37 ± 183 cell/mm3 e no grupo com resultados negativos foi 580 ± 2.890 cell/mm3 (p = 0,007). O limite superior aceitável de hemácias no LCR para não inibir o resultado do PCR foi 108 cells/mm3. As amostras de LCR com resultados negativos para RT-PCR EV tem mais eritrócitos em comparação com amostras com resultados positivos.

Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Cerebrospinal Fluid/cytology , Cerebrospinal Fluid/virology , Enterovirus/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction/methods , Erythrocytes , Reference Values , Time Factors , DNA, Viral/cerebrospinal fluid , RNA, Viral/cerebrospinal fluid , Sensitivity and Specificity , Enterovirus Infections/cerebrospinal fluid , Enterovirus Infections/virology , Erythrocyte Count , Meningitis, Viral/virology
Hist. ciênc. saúde-Manguinhos ; 22(1): 23-47, Jan-Mar/2015. graf
Article in English | LILACS, BDS | ID: lil-741507


Over recent years Brazil has played an increasingly active role internationally, the result of its model of integration and its foreign policy directives. The health sector is a valuable and strategic area for Brazilian technical cooperation to achieve various objectives, including its development goals. This article describes the main directives of Brazilian foreign policy, conceptually defining and characterizing South-South Cooperation, illustrated through an analysis of two Brazilian technical cooperation initiatives in healthcare: one in South America, the other in Africa. The study concludes that, irrespective of the interests and power asymmetries existing in South-South Cooperation, the objectives of this cooperation were achieved through the technical work.

Nos últimos anos, o Brasil foi ativo no âmbito internacional, tanto por seu modelo de inserção como pelas diretrizes de política externa. O setor saúde é uma ferramenta valiosa e estratégica utilizada pela cooperação técnica brasileira para lograr seus objetivos de desenvolvimento. Este artigo descreve as principais diretrizes de política externa brasileira, conceitua e caracteriza a Cooperação Sul-Sul, ilustrada mediante análise de duas iniciativas de cooperação técnica em saúde do Brasil: na América do Sul e na África. O estudo conclui que, independentemente dos interesses e das assimetrias de poder que existem na Cooperação Sul-Sul, os objetivos dessa cooperação foram alcançados por meio do trabalho técnico.

Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Disease Outbreaks , West Nile Fever/epidemiology , West Nile virus/isolation & purification , Age Factors , Antibodies, Viral/blood , Antibodies, Viral/cerebrospinal fluid , Cerebrospinal Fluid/immunology , Cerebrospinal Fluid/virology , Enzyme-Linked Immunosorbent Assay , Mortality , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , RNA, Viral/blood , RNA, Viral/cerebrospinal fluid , Survival Analysis , Serum/immunology , Serum/virology , Tunisia/epidemiology , West Nile Fever/pathology , West Nile Fever/virology
Article in English | WPRIM | ID: wpr-43987


BACKGROUND: Bacterial meningitis is an infectious disease with high rates of mortality and high frequency of severe sequelae. Early identification of causative bacterial and viral pathogens is important for prompt and proper treatment of meningitis and for prevention of life-threatening clinical outcomes. In the present study, we evaluated the value of the Seeplex Meningitis ACE Detection kit (Seegene Inc., Korea), a newly developed multiplex PCR kit employing dual priming oligonucleotide methods, for diagnosing acute meningitis. METHODS: Analytical sensitivity of the kit was studied using reference strains for each pathogen targeted by the kit, while it's analytical specificity was studied using the human genome DNA and 58 clinically well-identified reference strains. For clinical validation experiment, we used 27 control cerebrospinal fluid (CSF) samples and 78 clinical CSF samples collected from patients at the time of diagnosis of acute meningitis. RESULTS: The lower detection limits ranged from 101 copies/microL to 5x101 copies/microL for the 12 viral and bacterial pathogens targeted. No cross-reaction was observed. In the validation study, high detection rate of 56.4% was obtained. None of the control samples tested positive, i.e., false-positive results were absent. CONCLUSIONS: The Seeplex Meningitis ACE Detection kit showed high sensitivity, specificity, and detection rate for the identification of pathogens in clinical CSF samples. This kit may be useful for rapid identification of important acute meningitis-causing pathogens.

Acute Disease , Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Infant , Male , Meningitis/diagnosis , Middle Aged , Polymerase Chain Reaction , RNA, Bacterial/cerebrospinal fluid , RNA, Viral/cerebrospinal fluid , Reagent Kits, Diagnostic , Sensitivity and Specificity , Sequence Analysis, RNA
Rev. méd. Chile ; 139(12): 1588-1591, dic. 2011. ilus
Article in Spanish | LILACS | ID: lil-627593


Human herpesvirus 7 (HHV-7) may cause encephalomyelitis in immune competent adults. We report two patients infected by the virus. A 34-year-old male presenting with paraparesis and a sensitive deficiency located in D6 dermatome. Cerebrospinal fluid had 35 white blood cells per mm³ and 75 mg protein per dl. A PCR-microarray examination was positive for HHV-7. The patient was treated with prednisolone and ganciclovir with full recovery. A 27-year-old male presenting with headache, fever and diarrhea. Cerebrospinal fluid analysis showed 160 cells per mm³ and 75 mg protein per dl. Viral RNA detection was positive for HHV-7. The patient was managed with analgesia and rest and was discharged with the diagnosis of viral meningitis. Our communication supports the notion that HHV-7 may be considered as pathogen factor in humans, even in immune competent ones.

Adult , Humans , Male , Encephalitis, Herpes Simplex/virology , /isolation & purification , RNA, Viral/cerebrospinal fluid , Roseolovirus Infections , Diagnosis, Differential , Encephalitis, Herpes Simplex/cerebrospinal fluid , /genetics , Immunocompetence , Microarray Analysis/methods , Polymerase Chain Reaction , Roseolovirus Infections/cerebrospinal fluid
Rev. Inst. Med. Trop. Säo Paulo ; 53(4): 193-196, July.-Aug. 2011. tab
Article in English | LILACS | ID: lil-598598


The question of whether HIV-1 RNA in cerebrospinal fluid (CSF) is derived from viral replication in the central nervous system or simply reflects the transit of infected lymphocytes from the blood compartment has long been a matter of debate. Some studies found no correlation between CSF and plasma viral load, whereas others did. The lack of a correlation between the two compartments suggests that the presence of HIV-1 RNA is not simply due to the passive passage of the virus from blood to CSF but rather due to intrathecal replication. To evaluate the correlation between plasma and CSF HIV-1 RNA levels and to identify situations in which there is no correlation between the two compartments, seventy patients were prospectively studied. The association between CSF and plasma viral load was evaluated in the total population and in subgroups of patients with similar characteristics. A correlation between the CSF and plasma compartments was observed for patients undergoing highly active antiretroviral therapy (HAART), those with a CD4 T lymphocyte count lower than 200 cells/mm³, and those with increased CSF protein content. On the other hand, no correlation was observed for patients without adequate virological control, who had a CD4 count higher than 200 cells/mm³ and who did not use HAART. The correlation between the two compartments observed in some patients suggests that CSF HIV-1 RNA levels may reflect plasma levels in these subjects. In contrast, the lack of a correlation between the two compartments in patients who were not on HAART and who had normal CSF proteins and a poor virological control possibly indicates compartmentalization of the virus in CSF and, consequently, plasma-independent intrathecal viral replication.

Tem sido objeto de debate a questão se o RNA do HIV-1 no líquido cefalorraquidiano (LCR) é derivado da replicação viral no sistema nervoso central ou simplesmente reflete o tráfego de linfócitos infectados do compartimento sanguíneo. Alguns estudos não mostraram correlação entre a carga viral do plasma e LCR, mas outros sim. A falta de correlação entre os dois compartimentos sugere que a presença de RNA do HIV-1 não é simplesmente devido à passagem do vírus do plasma para o LCR, mas sim a uma replicação intratecal. Para avaliar a correlação entre os níveis de RNA do HIV-1 no plasma e no LCR e tentar identificar situações, na qual, não existe a correlação entre os dois compartimentos avaliaram-se setenta pacientes prospectivamente. A associação entre a carga viral do LCR e plasma foi avaliada na população total e em subgrupos de pacientes com características similares. A correlação entre os dois compartimentos foi observada em pacientes que estavam em uso da terapia antiretroviral (HAART), naqueles que tinham contagem de linfócitos CD4 menor que 200 céls/mm³ e naqueles com aumento da concentração de proteínas no LCR. Por outro lado, não houve correlação para os pacientes que não tinham um controle virológico adequado, os que tinham contagem de CD4 maior que 200 céls/mm³ e aqueles que não estavam usando HAART. A correlação entre os dois compartimentos observada em alguns pacientes sugere que os níveis de RNA do HIV-1 no LCR podem refletir os níveis plasmáticos nestes pacientes. E a falta de correlação ente os dois compartimentos em pacientes que não usavam HAART, nos que tinham uma concentração de proteínas no LCR normal, e nos que não apresentavam bom controle virológico, indica provavelmente a compartimentalização do vírus no LCR e consequentemente replicação viral intratecal independente da do plasma.

Adult , Female , Humans , Male , Middle Aged , Young Adult , AIDS-Related Opportunistic Infections/virology , Central Nervous System Viral Diseases/virology , HIV-1 , RNA, Viral/blood , RNA, Viral/cerebrospinal fluid , AIDS-Related Opportunistic Infections/blood , AIDS-Related Opportunistic Infections/cerebrospinal fluid , Antiretroviral Therapy, Highly Active , Central Nervous System Viral Diseases/blood , Central Nervous System Viral Diseases/cerebrospinal fluid , HIV-1 , Prospective Studies , Viral Load , Virus Replication
Arq. neuropsiquiatr ; 69(3): 475-481, June 2011. ilus, tab
Article in English | LILACS | ID: lil-592506


Viral meningitis is a common infectious disease of the central nervous system (CNS) that occurs worldwide. The aim of this study was to identify the etiologic agent of lymphomonocytary meningitis in Curitiba, PR, Brazil. During the period of July 2005 to December 2006, 460 cerebrospinal fluid (CSF) samples with lymphomonocytary meningitis were analyzed by PCR methodologies. Fifty nine (12.8 percent) samples were positive. Enteroviruses was present in 49 (83 percent) samples and herpes virus family in 10 (17 percent), of these 6 (10 percent) herpes simplex virus, 1 (2 percent) Epstein Barr virus, 2 (3 percent) human herpes virus type 6 and 1 (2 percent) mixed infection of enterovirus and Epstein Barr virus. As conclusion enterovirus was the most frequent virus, with circulation during summer and was observed with higher frequency between 4 to 17 years of age. PCR methodology is an important method for rapid detection of RNA enterovirus and DNA herpesvirus in CSF.

A meningite viral é uma síndrome infecciosa comum do sistema nervoso central (SNC), que ocorre no mundo inteiro. O objetivo deste estudo foi identificar o agente etiológico de meningite linfomonocitária em Curitiba, PR, Brasil. Durante o período de julho de 2005 a dezembro de 2006, 460 amostras com meningite linfomonocitária foram analisadas por metodologias de PCR. Cinquenta e nove (12,8 por cento) amostras foram positivas. Enterovirus estava presente em 49 (83 por cento) amostras e herpes vírus em 10 (17 por cento), destas 6 (10 por cento) HSV, 1 (2 por cento) EBV, 2 (3 por cento) HHV- 6 e 1 (2 por cento) infecção mista de enterovírus e EBV. Conclui-se que o enterovirus foi o vírus mais frequente, com a circulação durante o verão. Houve maior número de amostras positivas entre 4 a 17 anos. A metodologia de PCR é um importante método para a detecção rápida de RNA de enterovirus e DNA do herpesvirus no LCR.

Adolescent , Adult , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Young Adult , Enterovirus Infections/virology , Enterovirus/genetics , Herpesviridae Infections/virology , Herpesviridae/genetics , Meningitis, Viral/virology , Brazil , DNA, Viral/cerebrospinal fluid , Enterovirus Infections/diagnosis , Herpesviridae Infections/diagnosis , /genetics , /genetics , Meningitis, Viral/diagnosis , Polymerase Chain Reaction , RNA, Viral/cerebrospinal fluid , Simplexvirus/genetics
Arq. neuropsiquiatr ; 63(4): 907-913, dez. 2005. tab, graf
Article in English | LILACS | ID: lil-418994


INTRODUÇÃO: Os níveis de RNA do HIV-1 no plasma refletem a replicação viral sistêmica e a replicação no sistema nervoso central pode ocorrer independentemente da infecção sistêmica, mas a utilidade da medida destes níveis no líquido cefalorraqueano (LCR) permanece indefinida. OBJETIVO: Comparar os níveis de RNA do HIV-1 no LCR e plasma de pacientes sem doenças neurológicas e com diferentes doenças neurológicas, bem como correlacionar estes níveis com a sua evolução e o uso de antiretrovirais. MÉTODO: Foram avaliados 97 pacientes com suspeita de doença neurológica que realizaram punção lombar e que foram divididos em dois grupos: sem doenças neurológicas (23) e com doenças neurológicas (74). Metodologia NASBA foi usada para quantificação do RNA do HIV-1. RESULTADOS: A mediana da carga viral do LCR foi maior em pacientes com neurotoxoplasmose, neurocriptococose, demência pelo HIV e doença neurológica sem etiologia definida quando comparada aos pacientes sem doenças neurológicas. Não houve diferença da carga viral do plasma entre os pacientes com e sem doença neurológica. A mediana da carga viral do plasma e LCR foi maior nos pacientes que faleceram em relação aos tratados com sucesso. A carga viral do LCR e plasma foi menor nos pacientes com doenças oportunísticas que usavam HAART em relação aos que não a usavam. CONCLUSÃO: A carga viral no LRC foi maior nos pacientes com qualquer doença neurológica em relação aos sem doenças neurológicas, mas isto não ocorreu no plasma, sugerindo que doença neurológica influencia mais o compartimento do LCR que o do plasma, mas não foi possível diferenciar as doenças neurológicas pelos níveis de RNA do HIV-1 do LCR.

Adult , Female , Humans , Male , AIDS-Related Opportunistic Infections/virology , Central Nervous System Viral Diseases/virology , HIV-1 , RNA, Viral/blood , RNA, Viral/cerebrospinal fluid , AIDS-Related Opportunistic Infections/blood , AIDS-Related Opportunistic Infections/cerebrospinal fluid , Antiretroviral Therapy, Highly Active , Anti-HIV Agents/therapeutic use , Central Nervous System Viral Diseases/blood , Central Nervous System Viral Diseases/cerebrospinal fluid , HIV-1 , Prospective Studies , Statistics, Nonparametric , Viral Load , Virus Replication