Cerebrospinal fluid challenges for the diagnosis of herpes simplex infection in the central nervous system / Desafios do exame do líquido cefalorraquidiano para o diagnóstico de infecção por herpes simplex no sistema nervoso central

Abstract Herpes simplex virus (HSV) is a cause of a severe disease of the central nervous system (CNS) in humans. The demonstration of specific antibodies in the cerebrospinal fluid (CSF) may contribute to the retrospective neurological diagnosis. However, the commercial immunological tests for HSV infection are for use in serum samples. Objective: The aim of the present study was to adapt a commercial kit anti-HSV IgG used for serum samples to be performed with a CSF sample. Methods: Forty CSF specimens from 38 patients with suspected CNS HSV infection were serially diluted for detecting anti-HSV IgG by enzyme immunoassay (EIA). The same samples were also analyzed with the polymerase chain reaction (PCR). Results: The sensitivity of EIA test for HSV was 5% (dilution 1:40) and 65% (dilution 1:2) in CSF, and HSV DNA PCR was 15%. The combined analysis of EIA (dilution 1:2) and PCR increased the sensitivity up to 72.5%. The inflammatory CSF was associated with positive HSV PCR. Conclusions: We demonstrated the importance to adapt serological anti-HSV IgG EIA test for CSF assays to increase the accuracy of the analysis, considering the low concentration of specific antibodies in CSF.

Resumo O vírus herpes simples (HSV) é um dos agentes causadores de uma doença grave no sistema nervoso central (SNC) em humanos. A detecção de anticorpos específicos no líquido cefalorraquidiano (LCR) pode contribuir para o diagnóstico neurológico retrospectivo. Entretanto, os testes imunológicos comerciais são para uso em amostras de soro. Objetivo: Adaptar um kit comercial sorológico anti-HSV IgG para ser utilizado no de LCR. Metodos: Quarenta amostras de LCR de 38 pacientes com suspeita de infecção por HSV no SNC foram diluídas pesquisa de anticorpos anti-HSV IgG pelo método imunoenzimático (EIA). Além disso, as mesmas amostras também foram analisadas por reação em cadeia da polimerase (PCR). Resultados: A sensibilidade do teste EIA para o HSV consistiu em 5% (diluição 1:40) e 65% (diluição 1:2) no LCR, e o PCR do DNA do HSV, 15%. A análise combinada de EIA (diluição 1:2) e PCR aumentou a sensibilidade para 72,5%. Houve associação entre presença do LCR inflamatório e PCR positiva para HSV. Conclusões: Demonstramos a importância na adaptação previa do teste sorológico anti-HSV IgG EIA para ensaios do no LCR, a fim de aumentar a acuracia da análise, considerando a baixa concentração de anticorpos específicos no LCR.

FilmArray Meningitis/Encephalitis (ME) panel in the diagnosis of bacterial meningitis

ABSTRACT The precise diagnosis of bacterial meningitis is essential. Cytological and biochemical examination of cerebrospinal fluid (CSF) are not specific. Conventional methods for bacterial meningitis lack sensitivity or take too long for a final result. Therefore, other methods for rapid and accurate diagnosis of central nervous system infections are required. FilmArray meningitis/encephalitis (ME) panel is a PCR multiplex for simultaneous and rapid identification of 14 pathogens, including 6 bacteria, 7 viruses, and Cryptococcus. We evaluated 436 CSF samples submitted to FilmArray ME Panel. Among them, 25 cases were positive for bacteria, being Streptococcus pneumonia the most frequent (48 %). Among positive cases for bacteria, 60 % were positive only with FilmArray. All the bacterial meningitis cases in which the only positive test was FilmArray had CSF findings suggestive of bacterial meningitis, including neutrophilic pleocytosis, increased CSF protein and lactate, and decreased CSF glucose. These findings suggest that FilmArray may increase the diagnostic sensitivity for bacterial meningitis.

Parálisis facial neonatal: identificación del virus del herpes simple 1 en el líquido cefalorraquídeo. Caso clínico / Neonatal facial palsy: Identification of herpes simplex virus 1 in cerebrospinal fluid. Case report

En los neonatos, la parálisis facial es muy infrecuente y, por lo general, diagnosticada al nacer. Se presenta el primer caso de parálisis facial neonatal con identificación del virus del herpes simple 1 en el líquido cefalorraquídeo. Un varón de 35 días de vida acudió a Urgencias por la desviación de la comisura bucal hacia la izquierda y la ausencia de cierre del ojo derecho, sin sintomatología infecciosa ni antecedentes relevantes. La exploración física fue compatible con parálisis facial periférica. Las exploraciones complementarias de urgencia (hemograma, bioquímica, coagulación y citoquímica de líquido cefalorraquídeo) fueron normales. Fue ingresado con prednisolona oral y aciclovir intravenoso. La resonancia magnética craneal fue normal. A las 48 horas, se recibió el resultado positivo de la reacción en cadena de la polimerasa para el virus del herpes simple 1 en el líquido cefalorraquídeo. Con evolución favorable, completó 7 días de prednisolona oral y fue dado de alta tras 21 días de aciclovir intravenoso, con exploración neurológica previa normal.

Neonatal facial palsy is very uncommon and is generally diagnosed at birth. We present the first published case of neonatal facial palsy with identification of herpes simplex virus 1 in cerebrospinal fluid. A 35-day-old male was presented at the Emergency Department with mouth deviation to the left and impossibility of full closure of the right eye. There were no symptoms of infection or relevant medical history. Physical examination was compatible with peripheral facial palsy. Studies performed at admission were normal (blood count, biochemical analysis and coagulation blood tests and cerebrospinal fluid analysis). The patient was admitted on oral prednisolone and intravenous aciclovir. Cranial magnetic resonance was normal. Polymerase chain reaction test for herpes simplex virus 1 in cerebrospinal fluid was reported positive after 48 hours of admission. Patient followed good evolution and received prednisolone for 7 days and acyclovir for 21 days. At discharge, neurological examination was normal.

Encephalitis associated with Zika virus infection and reactivation of the varicella-zoster virus in a Brazilian child

Abstract We report a case of encephalitis associated with Zika virus infection and reactivation of varicella-zoster virus in the central nervous system of a Brazilian child. This case raises the possibility that reactivation of the latent varicella-zoster virus may be a mechanism of neurological impairment induced by acquired Zika virus infection.

Utilidad de la cuantificación de carga viral de VIH en líquido cefalorraquídeo, a propósito de un caso / Clinical utility of HIV viral load assessment in cerebral spinal fluid, a case report

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.

Presentación del síndrome de Wernicke-Korsakoff secundario a encefalitis por citomegalovirus, a propósito de un caso / Wernicke-Korsakoff syndrome secondary to cytomegalovirus encephalitis: A case report

Resumen El citomegalovirus (CMV) es uno de los microorganismos oportunistas con mayor prevalencia en pacientes inmunocomprometidos, aunque su reactivación ha descendido después de la introducción de la terapia antirretroviral altamente activa (Highly Active Antiretroviral Therapy, HAART). En las coinfecciones, la encefalitis se ha reportado como una de las condiciones más frecuentes. Se presenta el caso de un paciente adulto joven con infección por virus de la inmunodeficiencia humana (HIV) que tuvo un rápido deterioro neurológico evidenciado en síntomas y signos clínicos clásicos del síndrome de Wernicke-Korsakoff y que no presentaba factores de riesgo para deficiencia de tiamina. En las imágenes de la resonancia magnética cerebral, se detectaron hallazgos típicos del síndrome, y se identificó citomegalovirus (CMV) en el líquido cefalorraquídeo. Con el tratamiento específico para el CMV, se logró el control de los síntomas, aunque hubo secuelas neurológicas que mejoraron. Este es uno de los pocos casos reportados a nivel mundial de síndrome de Wernicke secundario a encefalitis por citomegalovirus.

Abstract Cytomegalovirus (CMV) is one of the opportunistic microorganisms with the highest prevalence in immunocompromised patients. Reactivation has decreased after the introduction of highly active antiretroviral therapy (HAART). Encephalitis has been reported in the coinfection as one of the most frequent presentations. We present the case of a young adult patient with HIV infection and rapid neurological deterioration due to classic clinical symptoms and signs of the Wernicke-Korsakoff syndrome, with no risk factors for thiamine deficiency, with images by nuclear magnetic resonance typical of the syndrome, and identification of cytomegalovirus in cerebrospinal fluid. The specific treatment for CMV managed to control the symptoms with neurological sequelae in progression towards improvement. This is one of the few cases reported in the literature of Wernicke syndrome secondary to cytomegalovirus encephalitis.

Red blood cells in cerebrospinal fluid as possible inhibitory factor for enterovirus RT-PCR / Presença de hemácias no líquido cefalorraquidiano como possível fator inibitório da RT-PCR para detecção de enterovirus

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.

Epidemiological surveillance of herpes viral encephalitis in Cordoba, Colombia / Vigilancia epidemiológica de encefalitis por herpes virus en Córdoba, Colombia

Objective To establish an epidemiological surveillance of viral herpes encephalitis in major hospitals of Monteria, Cordoba. Methods From September 2009 to December 2011, a descriptive study of cases of viral encephalitis was made in three hospitals in the city of Monteria. Cerebrospinal fluid (CSF) samples from 118 patients were included in the study. Clinical aspects, as well as cytochemical and microbiological analysis (Gram stain and culture) of CSF, were used for selecting the patients. Virus detection was performed by using multiplex nested PCR for Herpes simplex virus 1 and 2, Epstein Barr virus, Cytomegalovirus and Varicella zoster virus. Results Viral DNA of herpesvirus was detected in the CSFs of 30 (25.4 %) participants, as follows: 22 (18.6 %) Herpes simplex 1 and 2 viruses, 4 (3.3 %) Cytomegalovirus and 1 (0.8 %) Varicella zoster virus. Co-infections were observed in 3 patients (2.5 %), 1 case by HSV-VZV and 2 cases by CMV/HSV. The clinical manifestations of the patients included: headache (18.6 %), fever (14.4 %), asthenia (10.1 %), seizures (9.3 %), vomiting (8.4 %), and stiff neck (5.9 %). Thirty percent of the patients also had HIV-AIDS. A case fatality rate of 20 % was observed for the patients. Conclusions This paper shows that herpesvirus is a cause of infection of the CNS in patients from Cordoba. This study contributes to the epidemiology of encephalitis, as well as to patient management.(AU)

Objetivo Establecer una vigilancia epidemiológica de la encefalitis viral herpética en los principales hospitales de Montería, Córdoba. Materiales y Métodos Se realizó un estudio descriptivo de los casos de encefalitis viral entre septiembre de 2009 diciembre de 2011 en tres hospitales en la ciudad de Montería. Las muestras líquido cefalorraquídeo (LCR) de 118 pacientes fueron incluidos en el estudio. Los aspectos clínicos como el análisis citoquímico y microbiológico (tinción de Gram y cultivo) de LCR fueron utilizados para la selección de los pacientes. La detección de virus se realizó por PCR anidada multiplex para Herpes simplex virus 1 y 2, virus de Epstein Barr, virus zoster de la varicela y el citomegalovirus. Resultados Se detectó ADN viral del virus del herpes en 30 (25,4 %) muestras de LCR en los pacientes de la siguiente manera: 22 (18,6 %) Herpes simplex virus 1 y 2, 4 (3,3 %) Citomegalovirus y 1 (0,8 %) del virus de la varicela zóster. Se observaron Co-infecciones en 3 pacientes (2,5 %), 1 caso por el VHS-VZV y 2 casos por CMV / HSV. Las manifestaciones clínicas de los pacientes fueron: cefalea (18,6 %), fiebre (14,4 %), astenia (10,1 %), convulsiones (9,3 %), vómitos (8,4 %), y rigidez de nuca (5,9 %). El treinta por ciento de los pacientes también tenía VIH-SIDA. Se observó una tasa de letalidad del 20 % de los pacientes. Conclusiones Se demuestra que el herpesvirus es causa de infección del SNC en pacientes en Córdoba. Este estudio contribuye a la caracterización serológica viral epidemiológica de la encefalitis viral, así como en el manejo del paciente ya que se describen hallazgos clínicos importante en la población adulta estudiada.(AU)

Cerebrospinal fluid analysis in the HIV infection and compartmentalization of HIV in the central nervous system / Análise de LCR na infeção pelo HIV e compartimentalização do HIV no sistema nervoso central

The nervous system plays an important role in HIV infection. The purpose of this review is to discuss the indications for cerebrospinal fluid (CSF) analysis in HIV infection in clinical practice. CSF analysis in HIV infection is indicated for the diagnosis of opportunistic infections and co-infections, diagnosis of meningitis caused by HIV, quantification of HIV viral load, and analysis of CNS HIV compartmentalization. Although several CSF biomarkers have been investigated, none are clinically applicable. The capacity of HIV to generate genetic diversity, in association with the constitutional characteristics of the CNS, facilitates the generation of HIV quasispecies in the CNS that are distinct from HIV in the systemic circulation. CSF analysis has a well-defined and valuable role in the diagnosis of CNS infections in HIV/AIDS patients. Further research is necessary to establish a clinically applicable biomarker for the diagnosis of HIV-associated neurocognitive disorders.

O sistema nervoso representa um papel importante na infecção pelo HIV. O objetivo desta revisão é discutir as indicações para análise do líquido cefalorraquidiano (LCR) na infecção pelo HIV na prática clínica. A análise do LCR na infecção pelo HIV é indicada para o diagnóstico de infecções oportunistas e co-infecções, meningite pelo HIV, quantificação da carga viral de HIV e compartimentalização do HIV no SNC. Uma série de biomarcadores no LCR foi investigada, na literatura, porém não apresentam aplicabilidade clínica. A grande capacidade do HIV de gerar diversidade genética, associado a características constitucionais do SNC propicia o desenvolvimento quasiespécies distintas no SNC das circulantes sistemicamente. A análise do LCR na infecção pelo HIV é bem estabelecida no diagnóstico de infecções no CNS, contudo mais pesquisas é necessária para estabelecer a aplicabilidade clínica dos biomarcadores no diagnóstico de desordens cognitivas associadas ao HIV.

South-South Cooperation: Brazilian experiences in South America and Africa / Cooperação Sul-Sul: experiências brasileiras na América do Sul e na África

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.

Frecuencia de agentes virales en niños con afectación del sistema nervioso central en el estado Zulia, Venezuela: durante el año 2007 / Incidence of viral agents in children with central nervous system affectation in the state of Zulia, Venezuela: during 2007

Diversos virus afectan el sistema nervioso central (SNC) ocasionando encefalitis, principalmente en la edad pediátrica. Determinar la implicación de agentes virales en infecciones del sistema nervioso central (SNC) en niños del estado Zulia, Venezuela durante el año 2007. Se recolectaron 109 muestras de líquido cefalorraquídeo(LCR) y suero, provenientes de pacientes entre 1 día de nacido a 14 años, que presentaron sintomatología clínica sugestiva de afectación del SNC y cuyo estudio bacteriológico convencional de LCR resultó negativo. Se determinó la relación albúmina LCR/suero a fin de descartar contaminación, resultando 24 pares óptimos para la determinación por la técnica de ELISA de anticuerpos IgM específicos para los virus Herpes Simple (VHS), Epstein Barr (VEB), Dengue, Rubéola, Sarampión y Encefalitis Equina Venezolana (EEV). De los 24 casos analizados, 15 (62,5%) resultaron positivos. Los agentes causantes de encefalitis fueron: 11 casos de Dengue (45,8%) (p<0,05), 3 de VHS (12,5%) y un caso de VEB (4,2%). No se detectaron casos de Rubéola, Sarampión, ni EEV. La pleocitocis con predominio de linfocitos fue el hallazgo más frecuente en los casos con encefalitis viral (EV) confirmada, sin diferencias significativas al relacionarlo con el agente viral infectante. Se evidencia que una proporción significativa de los niños con encefalitis es debida a agentes virales y se destaca un incremento en los casos de dengue con afectación del SNC en la región

Various viruses affect the central nervous system (CNS) causing encephalitis, mainly in pediatric patients. To determine the involvement of viral agents for central nervous system (CNS) infections in children in the State of Zulia, Venezuela during the year 2007. 109 samples of cerebrospinal fluid (CSF) and serum were collected from patients between 1 day and 14 years of age, who presented clinical symptoms suggestive of CNS involvement and whose conventional CSF bacteriological study proved negative. The CSF Albumin /serum relation ship was determined in order to rule out contamination, resulting in 24 optimal pairs for determining the IgM and IgG antibodies specific for herpes simplex virus (HSV), Epstein Barr (EBV), Dengue fever, rubella, measles and Venezuelan Equine Encephalitis (VEE) using the ELISA technique. Of the 24 cases examined, 15 (62.5%) were positive. The causative agents for encephalitis were 11 cases of Dengue (45.8%) (p<0.05), 3 VHS (12.5%) and 1 case of EBV (4.2%). There were no cases of rubella, measles or VEE. Pleocytosis with lymphocyte predominance was the most common finding in cases with confirmed viral encephalitis (VE), without significant differences related to the infecting viral agent. Results show that a significant proportion of encephalitis in children is due to viral agents, highlighting an increase in dengue cases with CNS affection in the region

Análise molecular de patógenos pela reação em cadeia da polimerase no líquido cefalorraquiano de pacientes infectados com HIV / Molecular analysis of pathogens in cerebrospinal fluid by the polymerase chain reaction in HIV-infected patients

Introdução: A reação em cadeia da polimerase (PCR) foi teste de grande impacto no diagnóstico das meningites e encefalites linfocíticas durante a última década. Esse método foi extensivamente usado no diagnóstico das infecções do sistema nervoso central (SNC), devido a sua habilidade em detectar amostras mínimas de DNA-alvo no líquido cefalorraquiano. Objetivo: O objetivo deste estudo foi identificar a prevalência dos patógenos oportunistas responsáveis por causar problemas neurológicos em pacientes infectados com o vírus da imunodeficiência humana (HIV) e avaliar sua associação com os achados clínicos, laboratoriais e da tomografia computadorizada cerebral (TCC). Pacientes e métodos: Um estudo transversal foi realizado em 203 amostras de líquido cefalorraquiano (LCR) de pacientes do sul do Brasil infectados com HIV e com aparente encefalite e meningite linfocíticas. As amostras foram analisadas para os seguintes agentes pelo método da reação em cadeia da polimerase "nested" ou dupla (N-PCR): citomegalovírus, vírus do Epstein-Barr, vírus do herpes simplex tipos 1 e 2, vírus da varicella zoster, vírus do herpes humano tipo 6, vírus JC, Toxoplasma gondii e micobactérias. Resultado: Pelo menos um patógeno foi encontrado em 77 (38%) dos indivíduos. O Epstein-Barr foi o mais prevalente, com 40 casos (19,7%), seguido pelo citomegalovívus, com 12 casos (15%) e pelo vírus JC, em 9 casos (4,4%). Um N-PCR positivo mostrou associação com aumento de proteínas e de celularidade (P=0,001), meningismo (P=0,017) e tomografia computadorizada anormal (P=0,006). Conclusão: O painel de PCR empregado foi efetivo na identificação de infecções neurológicas severas em pacientes HIV positivos (AU)

Introduction: Polymerase chain reaction (PCR) has had great impact on the diagnosis of lymphocytic meningitis and encephalitis over the last decade. It has been extensively used in the diagnosis of central nervous system (CNS) infections for its ability to detect small amounts of target DNA in the cerebrospinal fluid (CSF). Objective: The aim of this study was to identify the prevalence of opportunistic pathogens responsible for neurological disorders in patients infected with human immunodeficiency virus (HIV) and to evaluate its association with clinical, laboratory and cerebral computed tomography (CCT) findings. Patients and methods: A cross-sectional study was performed on 203 cerebrospinal fluids (CSF) from HIV-infected patients from Southern Brazil, with apparent lymphocytic meningitis and encephalitis. CSF samples were analyzed with probes for cytomegalovirus, Epstein-Barr virus, herpes simplex virus types 1 and 2, varicella zoster virus, human herpes virus type 6, JC virus, Toxoplasma gondii and mycobacterium in nested polymerase chain reaction (N-PCR). Results: At least one pathogen was found in 77 (38.0%) individuals. Epstein-Barr virus was the most prevalent with 40 cases (19.7%), followed by cytomegalovirus with 12 cases (5.9%) and JC virus with 9 cases (4.4%). Positive NPCR showed association with high spinal fluid protein and cell count (P=0.001), meningism (P=0.017) and abnormal CCT (P=0.006). Conclusion: The PCR panel used was effective in screening several neurological infections in HIV positive patients (AU)

case of herpes simplex encephalitis with unusual presentation

History and investigation are the leader in helping physicians to reach a proper diagnosis; this aim is applied in both simple and complicated cases such as Herpes Simplex Encephalitis [HSE]. USE can be consider as life threatening condition due to its high mortality rate in delayed treated condition and neurological sequelae even if treatment initiated early. This was applied on a four years old girl presented to us with one day history of head trauma which was supported by hemorrhage seen in CT-Head, 72 hours later her condition deteriorate by recurrent convulsions, aphasia, change level of consciousness and spikes of fever, diagnosis of USE was delayed till day 10 of presentation when she start to have frequent attacks of convulsions, which was proved by positive CSF PCR for HSV

Biología molecular en infectología: parte II, diagnóstico molecular de agentes infecciosos / Molecular biology in infectius diseases: part II molecular diagnosis of infectious agents

Las aplicaciones diagnósticas de la biología molecular para enfermedades infecciosas son extremadamente variadas y aplicables a cualquier problema diagnóstico. En agentes virales de la familia Hepersviridae, los más usados se basan en la amplificación del gen de la enzima ADN polimerasa que permite la detección de virus herpes simplex (VHS) 1 y 2, virus varicela-zoster (VVZ), citomegalovirus (CMV), virus de Epstein Barr (VEB) y herpesvirus humano (HVH) 6 en forma simultánea. Esta metodología ha detectado la coinfección por VHS 1 y VZV en muestras de líquido cefalorraquideo. En CMV son utilizados en el monitoreo de la reactivación de CMV en pacientes inmunosuprimidos siendo capaz de detectar reactivación viral con una semana de anticipación a la aparición de los síntomas. Los métodos moleculares han permitido la identificación del VEB en una proporción de 8 a 20 por ciento de casos de cáncer gástrico los cuales poseen una cepa única a pesar de la presencia de múltiples cepas en la población sana. Estas asociaciones entre virus y cáncer también se han descrito para el virus papiloma humano y cáncer pulmonar. En agentes bacterianos, la detección y cuantificación de Bordetella pertussis es otra aplicación relevante ya que podría convertirse en un método de diagnóstico rápido y predictivo de severidad de enfermedad en niños menores de 6 meses. La caracterización de cepas de Helicobacter pylori en relación con cáncer gástrico y enfermedadulcerosa péptica, y la caracterización de cepas nosocomiales de Staphylococcus aureus resistente a meticilina (SAMR), son ejemplos de las potencialidades de los métodos moleculares en la tipificación de microorganismos. En el diagnóstico de agentes causantes de patologías respiratorias como Mycobacterium tuberculosis, Pneumocystis carinil y agentes atipicos de infecciones respiratorias, estos métodos han permitidoel diagnóstico a partir de muestras de obtención no invasora. Finalmente, también han demostrado su aporto en el diagnóstico de infecciones micóticas (candidiasis y aspergilosis), n particular en pacientes inmunocomprometidos

Detección de enterovirus mediante transcripción reversa y reacción de polimerasa en cadena en líquido cafalorraquídeo de niños con meningitis aséptica / Detection of enterovirus by reverse transcriptase and polymerase chain reaction in cerebrospinal fluid samples of children with aseptic meningitis

Los enterovirus causan cerca de 90 por ciento de las meningitis asépticas (MA). La ausencia de un buen método de diagnótico etiológico determina con frecuencia tratamientos innecesarios. En este estudio se comparó la detección de enterovirus en líquido cefalorraquídeo (LCR) usando cultivos celulares e inmunofluorescencia indirecta (CC/IFI), con la reacción de polimerasa en cadena (RT-PCR). Se estudiaron 22 muestras de LCR provenientes de niños con MA (casos) y 17 de niños con otras patologías (controles). Todos fueron atendidos en servicios pediátricos del Hospital Carlos Van Buren de Valparaiso. Hubo 100 por ciento de concordancia entre ambos métodos en los 22 niños con MA, encontrándose 21 casos positivos para enterovirus. Entre los 17 controles, 2 muestras resultaron por RT-PCR y una de ellas lo fue también por CC-IFI. La RT-PCR resultó un método sencible y específico. Considerando la rapidez con que se puede tener los resultados es esperable que su uso rutinario sea beneficioso para los pacientes con MA

Polymerase chain reaction for the laboratory diagnosis of aseptic meningitis and encephalitis

A protocol for testing cerebrospinal fluid specimens using a range of PCR assays for the diagnosis of central nervous system infection was developed and used to test prospectively 383 specimens. PCR assays were used for the detection of adenovirus, Borrelia burgdorferi, enteroviruses, Epstein Barr virus, cytomegalovirus, herpes simplex virus, human herpes virus type 6, JC virus, Leptospira interrogans, Listeria monocytogenes, lymphocytic choriomeningitis virus, measles virus, mumps virus, Mycobacterium sp., Mycoplasma pneumoniae, Toxoplasma gondii and varicella zoster virus. Of the 383 specimens tested in this study, 46 (12.0 percent) were found to be positive. The microorganisms detected were CMV, enterovirus, Epstein Barr virus, herpes simplex virus, human herpes virus type 6, JC virus, L. monocytogenes, Mycobacterium genus, Toxoplasma gondii and varicella zoster virus. The introduction of the PCR protocol described has improved the diagnosis of a range of central nervous system infections in our laboratory. We believe however that further evaluation of these assays in immunocompromised patients is necessary to better determine the predictive value of positive PCR results in these patient groups

IGA1 proteases: ocorrência em agentes estiológicos de meningite / IgA1 proteases: ocurrence in meningitis etiological agents

Dentre os fatores de virulência em bactérias investigados na Seçäo de Bacteriologia do Instituto Adolfo Lutz, tem sido avaliada a ocorrência de proteases à imunoglobulina A (IgA1 proteases) em agentes etiológicos de meningite. Estas enzimas extracelulares säo produzidas por diferentes patógenos de mucosa, incluindo os três principais agentes das meningites bacterianas: Neisseria menigitidis, Haemophilus influenzae e Streptococus pneumoniae. IgA1 proteses foram detectadas por imunoeletroforese em 28 de 39 cepas estudadas, bactérias e leveduras. Com exceçäo de 7 bactérias obtidas de coleçäo de cultura, todos os microrganismos foram isolados de líquido cefalorraquidiano. Nos três agentes etiológicos mais frequentes de meningite foi observada a presença de IgA1 proteases, como também em quatro outras bactérias, das quais duas de coleçäo, e näo citadas na literatura como produtoras destas enzimas

Morphometric analysis of McCoy cells inoculated with cerebrospinal fluid from patients with rabies

To demonstrate the potential of McCoy cells for the isolation of rabies virus from the cerebrospinal (CSF) fluid of a patient with a diagnosis of rabies, McCoy cells were inoculated with CSF from a patient with a clinical diagnosis of rabies and investigated in terms of morphometric aspect using the JAVA analysis system for the quantification of the increased size of infected cells compared to noninfected cells. The cells were also examined in terms of specific staining for the diagnosis of rabies by the method of Sellers for the observation of intracytoplasmic inclusions and by specific immunofluorescence staining for rabies virus. Infected cells showed changes in cell permeability and morphologic modifications which differed significantly compared to normal cells (p<0.001) when analysed by the Mann-Whitney and Kruskal-Wallis tests. Intense activity of the endoplasmic reticulum was also observed, as indicated by the presence of intracytoplasmic inclusions visualized by specific staining. The present study demonstrated the isolation of rabies virus from the CSF of a patient with rabies, showing that McCoy cells can be used for the laboratory diagnosis of patients suspected to have rabies.

Meningitis aséptica y por enterovirus en el niño mayor / Aseptic meningitis and enterovirus meningitis in children

Durante el año 1993 se presentó un aumento de 3 veces el número promedio de casos de meningitis asépticas en el Hospital Roberto del Río. Se presentan las características clínicas de 50 niños mayores con meningitis aséptica, y se detallan las de 16 casos de ellos con diagnóstico comprobado de meningitis por enterovirus. El cuadro clínico fue benigno, no hubo complicaciones, muerte o secuelas al alta. Dentro de los hallazgos de laboratorio, se destaca el promedio de neutrófilos en el 30 por ciento de los casos en la punción lumbar de control,luego de 24 horas o más de evolución de la enfermedad.El echovirus 30 fue el enterovirus predominante. Se comparan los hallazgos con la literatura, y se comenta la utilidad de una segunda punción lumbar en casos como éstos