Inmunofluorescencia indirecta versus reacción de polimerasa en cadena para el diagnóstico de virus respiratorios en niños ingresados en un hospital de la Región Metropolitana / Indirect immunofluorescence technique versus polymerase chain reaction for the diagnosis of respiratory viruses in children admitted to a hospital in the Metropolitan Region

Resumen Introducción: La temprana detección viral en infecciones respiratorias agudas (IRA) es esencial para establecer una terapia apropiada y prevenir el contagio intrahospitalario. Objetivo: Comparar la eficacia de la técnica de inmunofluorescencia indirecta (IFI) con la reacción de polimerasa en cadena (RPC) para identificar virus respiratorios en niños hospitalizados por IRA. Métodos: Se incluyeron 47 aspirados nasofaríngeos de niños ≤ 2 años con IRA. La IFI incluyó virus respiratorio sincicial (VRS), adenovirus, influenza A y B y parainfluenza. La RPC incluyó, además, la detección de metapneumovirus, enterovirus/rinovirus, bocavirus y coronavirus. Se estimó sensibilidad, especificidad, valor predictor positivo y negativo (VPP/VPN) y correlación kappa para VRS mediante IFI en comparación a la RPC. Resultados: La IFI detectó únicamente VRS (29; 61,7%). La RPC detectó diversos virus, entre ellos VRS en 26 casos (55,3%), seguido por bocavirus (29,8%), enterovirus/ rinovirus (21,3%), adenovirus (14,9%) y parainfluenza (4,3%) entre otros, con 35,5% de co-infección. La IFI presentó sensibilidad: 85,7%, especificidad: 73,6%, VPP: 82,7%, VPN: 77,7% y kappa: 0,5990 (IC 95%; 0,36360,8346) para VRS. Conclusión: La IFI presenta buena sensibilidad, pero moderada especificidad para VRS. Sin embargo, falla en la detección de otros virus respiratorios. La introducción de RPC permitiría mejorar el diagnóstico etiológico de las IRA de origen viral.

Background: Early viral detection in acute respiratory infections (ARI) is essential to establish appropriate therapy and prevent nosocomial transmission. Objective: To compare the efficacy of indirect immunofluorescence technique (IIF) with the polymerase chain reaction (PCR) to identify respiratory viruses in children hospitalized for ARI. Methods: 47 nasopharyngeal aspirates of children ≤ 2 years with ARI were included. IFI included respiratory syncytial virus (RSV), adenovirus, influenza A and B and parainfluenza. PCR also included the detection of metapneumovirus, enterovirus/rhinovirus, bocavirus and coronavirus. Sensitivity, specificity, positive and negative predictive value (VPP/NPV) and kappa correlation for RSV were estimated by IIF compared to PCR. Results: The IIF detected only RSV (29; 61.7%). PCR detected several viruses, including RSV in 26 cases (55.3%), followed by bocavirus (29.8%), rhinovirus/enterovirus (21.3%), adenovirus (14.9%) and parainfluenza (4,3%) among others, with 35.5% of coinfection. The IIF presented sensitivity: 85.7%, specificity: 73.6%, PPV: 82.7%, NPV: 77.7% and kappa: 0.5990 (95% CI, 0.3636-0.8346) for RSV. Conclusion: The IIF presents good sensitivity, but moderate specificity for RSV. However, IIF fails to detect other respiratory viruses. The introduction of PCR would improve the etiological diagnosis of ARI of viral origin.

Assessment of enteric viruses in a sewage treatment plant located in Porto Alegre, southern Brazil / Avaliação de vírus entéricos em uma estação de tratamento de esgoto localizada em Porto Alegre, sul do Brasil

In order to verify the microbial quality of the influents and effluents of one STP from southern Brazil, an eight-month survey was conducted to examine the presence of total and fecal coliforms and of adenovirus (HAdV), enterovirus (EV), genogroup A rotaviruses (GARV) and Torque teno virus (TTV), in treated effluent samples from São João/Navegantes STP, Porto Alegre (Brazil). A total of 16 samples were collected, eight of influent (raw sewage, prior to treatment), and the other eight of the effluent (post-treatment sewage). Total and fecal coliform levels ranging from 3.6 × 10(4) to 4.4 × 10(7) MPN/100 mL and 2.9 × 10³ to 1.7 × 10(7) MPN/100 mL, were detected in all samples. In raw sewage, HAdV (25%) and GARV (28.6%) viral genomes were detected. The analysis of effluent samples revealed the presence of HAdV (50%), EV (37.5%), and TTV (12.5%) genomic fragments. All samples, regardless of the month analysed, presented detection of a least one virus genus, except for in April. Higher virus detection rates were observed in treated sewage samples (62.5%), and in 80% of them (effluent positive samples) HAdV was detected. Results showed that improvements in sewage monitoring and treatment processes are necessary to reduce the viral and bacterial load on the environment in southern Brazil. To the knowledge of the authors, this is the first study showing the monitoring of viral genomes in influent and effluent samples from a STP located in Porto Alegre (Rio Grande do Sul, Brazil), southern Brazil.

Com o intuito de verificar a qualidade microbiológica de afluentes e efluentes de uma estação de tratamento de esgoto (ETE), um monitoramento de oito meses foi realizado para examinar a presença de coliformes totais e fecais, e de adenovírus (HAdV), enterovírus (EV), rotavírus do genogrupo A (GARV) e torque teno vírus (TTV), em amostras de esgoto tratado da ETE São João/Navegantes, em Porto Alegre-RS, Brasil. Um total de 16 amostras foi coletado, sendo oito de afluente (esgoto bruto, anterior ao tratamento) e oito de efluente (esgoto tratado). Os níveis de coliformes totais e fecais variaram entre 3,6 × 10(4) e 4,4 × 10(7) MPN/100 mL e 2,9 × 10³ e 1,7 × 10(7) MPN/100 mL, respectivamente, tendo sido estes detectados em todas as amostras. No esgoto bruto, foram detectados os genomas virais de HAdV (25%) e GARV (28,6%). A análise das amostras de efluente revelou a presença de fragmentos genômicos de HAdV (50%), EV (37,5%) e TTV (12,5%). Todas as amostras, independentemente do mês analisado, possibilitaram a detecção de pelo menos um gênero viral, exceto no mês de abril. Altas taxas de detecção viral foram observadas em amostras de esgoto tratado (62,5%), sendo que o HAdV foi detectado em 80% dessas amostras de efluente positivas. Os resultados mostram que aprimoramentos no processo de tratamento e monitoramento do esgoto são necessários para reduzir a carga viral e bacteriológica no ambiente do Sul do Brasil. Ao conhecimento dos autores, este é o primeiro estudo de monitoramento de genomas virais em amostras de afluente e efluente de uma ETE localizada em Porto Alegre-Rio Grande do Sul, Brasil.

Acute respiratory viral infections in children in Rio de Janeiro and Teresópolis, Brazil / Infecções respiratórias agudas causadas por vírus em crianças do Rio de Janeiro e de Teresópolis, Brasil

The frequency of viral pathogens causing respiratory infections in children in the cities of Rio de Janeiro and Teresópolis was investigated. Nasal swabs from children with acute respiratory illnesses were collected between March 2006 and October 2007. Specimens were tested for viral detection by conventional (RT)-PCR and/or real time PCR. Of the 205 nasal swabs tested, 64 (31.2%) were positive for at least one of the viral pathogens. Single infections were detected in 56 samples, 50 of those were caused by RNA viruses: 33 samples tested positive for rhinovirus, five for influenza A, five for metapneumovirus, four for coronavirus and, three for respiratory syncytial virus. For the DNA viruses, five samples were positive for bocavirus and one for adenovirus. Co-infections with these viruses were detected in eight samples. Our data demonstrate a high frequency of viral respiratory infections, emphasizing the need for a more accurate diagnosis particularly for the emerging respiratory viruses. The fact that the emerging respiratory viruses were present in 9.2% of the tested samples suggests that these viruses could be important respiratory pathogens in the country.

Neste estudo foi investigada a frequência de patógenos virais causando infecção em crianças nas cidades do Rio de Janeiro e Teresópolis. Foram coletados 205 swabs nasais de crianças com infecção aguda do trato respiratório no período de março de 2006 a outubro de 2007. Os espécimes foram testados para detecção de vírus através de (RT)-PCR e/ou PCR em tempo real. Dentre as 205 amostras testadas, 64 (31,2%) foram positivas para pelo menos um vírus. Infecções causadas por um único agente viral foram detectadas em 56 amostras, 50 das quais eram causadas por vírus de RNA: 33 amostras foram positivas para rinovírus, cinco amostras foram positivas para influenza A, cinco amostras foram positivas para metapneumovírus, quatro amostras foram positivas para coronavírus e três amostras foram positivas para vírus respiratório sincicial. Para os vírus de DNA foram detectadas cinco amostras positivas para bocavírus humano e uma amostra positiva para adenovírus. Foram identificados oito casos de co-infecção. Nossos dados demonstram frequência elevada de infecções respiratórias virais, enfatizando a necessidade de um diagnóstico mais acurado destes patógenos, principalmente os vírus considerados emergentes. O fato de alguns vírus respiratórios emergentes terem sido detectados em 9,2% das amostras testadas sugere que estes vírus podem ser patógenos respiratórios importantes no país.

Viral acute gastroenteritis: clinical and epidemiological features of co-infected patients

BACKGROUND: Acute gastroenteritis (AGE) is a common disorder that affects children worldwide. It is usually caused by viral agents, including rotavirus, enteric adenovirus, norovirus, and astrovirus groups. Currently, there are few reports about co-infection among these viruses, mainly in Brazil. METHODS: This is a retrospective study in which 84 rotavirus-positive samples from hospitalized patients at a teaching hospital in Southern Brazil, collected in the 2001-2010 period, were analyzed by polymerase chain reaction (PCR) and reverse transcription - polymerase chain reaction (RT-PCR), for the investigation of enteric adenovirus, astrovirus, and norovirus. RESULTS: In total, 12 of the 84 (14%) samples were positive to enteric adenovirus or norovirus. Clinical, laboratory, and demographic data showed statistically significant differences between mono and co-infected patients, including age and depletion rate. CONCLUSIONS: These findings highlight the need for implementation of other enteric virus detection assays in clinical diagnosis for a complete laboratory investigation of hospitalized pediatric patients with AGE, in order to understand the impact of these pathogens on disease severity, spread within hospital, and consequently, prevent the dissemination of nosocomial infections.

Laboratory results in ocular viral diseases: implications in clinical-laboratory correlation

PURPOSE: To document etiology and predictive value of clinical diagnosis in laboratory confirmed viral diseases. METHODS: Reports of culture-positive cases of samples collected from patients presenting from January 1987 - December 2001 were evaluated. RESULTS: One thousand nine hundred and sixty-four (1964) cultures were submitted during 1987-2001. Twenty-six percent were positive (514). Human herpesvirus 1 was the most frequent agent isolated from all positive culture (56 percent). Adenovirus was the most common virus isolated from conjunctiva (66 percent), human herpesvirus 1 from lid and cornea (76 percent, 88 percent) and cytomegalovirus from vitreous (27 percent). Some unusual pathogens were recovered from conjunctiva as cytomegalovirus and from cornea as adenovirus, enterovirus and cytomegalovirus. Recognition of common viral syndromes was human herpesvirus 1 (88 percent), epidemic keratoconjunctivitis (88 percent), acute hemorrhagic conjunctivitis (70 percent) and varicella zoster virus (100 percent). However, some misdiagnosed cases were observed. Thirteen percent of conjunctivitis thought to be caused by herpes were due to adenovirus, 3.2 percent to Enterovirus, 3.2 percent to varicella zoster virus and 3.2 percent to human cytomegalovirus. Also, 5 percent of cases with a clinical diagnosis of herpes keratitis were caused by adenovirus and 2.7 percent by enterovirus. Finally, 4.8 percent of cases thought to be adenovirus conjunctivitis were herpes conjunctivitis. CONCLUSIONS: Human herpesvirus 1 remains the most frequently isolated virus from ocular sites in general (56 percent). Nonherpetic corneal isolates were in decreasing order: adenovirus, enterovirus and cytomegalovirus. Clinical and laboratory correlation was less than 90 percent. The most misdiagnosed cases were herpes conjunctivitis and keratitis, some cases of adenovirus conjunctivitis some cases of acute hemorrhagic conjunctivitis. It is essential that a rapid and...

OBJETIVOS: Documentar a etiologia e prever a importância do diagnóstico clínico em doenças virais oculares confirmadas em laboratório. MÉTODOS: Todos os relatórios de pacientes com cultura viral positiva durante o período de janeiro 1987 - dezembro 2001 foram analisados. RESULTADOS: Quinhentos e quatorze (514) casos foram encontrados. Em geral, herpesvírus 1 humano foi o agente mais freqüentemente isolado. Adenovírus foi o vírus mais comumente isolado da conjuntiva (66 por cento), herpesvírus 1 humano das pálpebras e córnea (76 por cento, 88 por cento respectivamente) e citomegalovírus do vítreo (27 por cento). Alguns agentes não usuais foram isolados da conjuntiva como citomegalovírus e da córnea como adenovírus, enterovírus e citomegalovírus. Reconhecimento das síndromes virais comuns foi: herpervírus 1 humano (88 por cento), ceratoconjuntivite epidêmica (88 por cento), conjuntivite aguda hemorrágica (70 por cento). Porém, alguns casos com diagnóstico incorreto foram observados. Treze por cento das conjuntivites com diagnóstico de herpes foram causadas por adenovírus, 3,2 por cento por enterovírus, 3,2 por cento por varicella zoster vírus e 3,2 por cento por citomegalovírus. Também, 5 por cento dos casos com diagnóstico clínico de ceratite herpética eram adenovírus e 2,7 por cento enterovírus. Além disso, 4,8 por cento dos casos em que se pensaram em conjuntivite por adenovírus, eram conjuntivite por herpes. Finalmente, 30 por cento dos casos em que se diagnosticaram conjuntive hemorrágica aguda, o agente etiológico era adenovírus. CONCLUSÃO: Em geral herpesvírus humano 1 continua a ser o vírus mais comum encontrado nas infecções oculares (56 por cento). Agentes não herpéticos isolados da córnea foram em ordem decrescente: adenovírus, enterovírus e citomegalovírus. A correlação entre o diagnóstico clínico e laboratorial foi menor do que 90 por cento. Um diagnóstico rápido e específico é essencial em casos de apresentações virais atípicas...

Gastritis plasmocelular crónica activa asociada a citomegalovirus / Chronic active plasmacytic gastritis associated with cytomegalovirus

La gastritis plasmocelular crónica activa (GPCA) se caracteriza por la presencia de infiltrados inflamatorios crónicos predominantemente plasmocelulares que comprometen las células del cuello de las glándulas gástricas. Tanto la GPCA como la gastropatía de Ménétrier han sido vinculadas con el citomegalovirus (CMV). Con el propósito de probas esta asociación, se evaluó la relación fovéolo-glandular (F/G) y se determinó la presencia del AND (ácido desoxirribonucleico) del CMV mediante PCR (reacción en cadena de polimerasa) en 12 casos y 13 controles. Los ejemplos de GPCA presentaron enfermedad de Ménétrier en 2 casos; hiperplasia foveolar, en 6 casos, y relación F/G conservada, en 3 casos. En ninguno visualizaron imágenes de gastritis linfocitaria, ni alteraciones citopáticas por CMV. Tres casos se asociaron con carcinoma gástrico. La relación F/G fue menor de 1 en todos los controles. Once de los 12 casos con GPCA mostraron amplificación para le AND del CMV y los 13 controles fueron negativos. Los hallazgos sugieren una fuerte asociación, posiblemente secuencial, entre la infección por el CMV, la GPCA, la hiperplasia foveolar (con o sin desarrollo de enfermedad de Ménétrier) y el carcinoma gástrico. La GPCA podría representar un marcador histológico de la presencia de CMV en la zona germinativa de los cuellos glandulares. La propuesta es paralela a los hallazgos, las lesiones y la secuencia observables en la infección hepática por VHB (irus de hepatitis B), con el desarrollo de hepatitis crónica, cirrosis y hepatocarcinoma.

Virus TT (TTV) ¿Es un verdadero causante de hepatis? / Is TT virus (TTV) a true hepatitis virus cause?

El virus TT (TTV), se detectó en sangre de tres pacientes con hepatitis postransfusional que eran negativos para todos los marcadores de virus de hepatitis conocidos. Los títulos de ADN viral estaban correlacionados con los niveles de aminotransferas séricas (ALT). Además se aisló el virus de hígado con títulos iguales o mayores a los sueros correspondientes. Posteriormente se comprobó que la mayoría de las personas TTV positivas no presentan evidencias bioquímicas o histológicas de enfermedad hepática. TTV tiene una distribución mundial, con una alta prevalencia en la población general, y existen evidencias de que es transmisible a través de transfusiones sanguíneas, por vía entérica y de madre a hijo. Existe una importante cantidad de estudios que sugieren que este virus no es una causa significativa de enfermedad hepática crónica o aguda y hasta ahora no se ha descripto ninguna asociación de la infección con otras enfermedades.

Clearance of hepatitis TT virus infection among thalassemia children and IVDU.

The novel transfusion transmissible hepatitis virus TTV first isolated by a group from Japan has predominantly been detected in members of groups at high risk for contracting blood borne viruses. Aside from elevated liver enzymes, the symptoms associated with its infection have been reported to range from asymptomatic to hepatic failure. The purpose of the present study was to determine if and to what extent the host's immune response is capable of clearing TTV infection. Hence, we extracted DNA from sera obtained from altogether 201 intravenous drug users (IVDU) and 80 thalassemia children--both groups at high risk of parenteral exposure--and performed PCR using semi-nested primers. Those positive for TTV DNA were once again subjected to PCR after approximately one year in order to determine how many still harbored the virus. Our results showed TTV DNA to be absent in merely 20.6% of the formerly positive IVDU, whereas it was still present in all the thalassemia children who could be tested for the second time. Based on the small sample size and the high-risk environment, these results ought to be interpreted with caution and definitely merit further investigation.

Ausência do HBV-DNA por PCR em soros de doadores de sangue com HBsAg negativo e anti-HBc positivo em regiäo de alta prevalência de hepatite B / Absence of HBV-DNA by PCR in blood donor sera with negative HBsAg and positive anti-HBc from a population with high prevalence of Hepatitis B

Foram estudados soros de 50 doadores voluntários de sangue, com antígeno de superfície da hepatite B (HBsAg) negativo, anticorpo do antígeno core da hepatite B (anti-HBc) positivo e níveis normais de alanina aminotransferase (ALT), provenientes de uma populaçäo de alta prevalência de hepatite B.As estatísticasdo HEMEPAR (Instituto de Saúde do Paraná - Centro de Hematologia e Hemoterapia do Paraná), regional de Cascavel mostram uma prevalência de anti-HBc positivo em 1995 e 1996 de 33,8 porcento e 31,1 porcento na regiäo Oeste do Paraná. Um método sensível, nested-PCR foi usado para pesquisar o DNA viral da hepatite B (HBV-DNA). O limite mínimo de detecçäo foi de 300 genomas por mL. Em todos os 50 soros anti-HBc positivo e HBsAg negativo analisados pela técnica de Reaçäo em Cadeia pela Polimerase näo foi encontrado DNA viral da hepatite B (HBV-DNA). Estes achados indicam que o descarte de todas as unidades de sangue ou hemoderivados com os marcadores, anti-HBc positivo e HBsAg negativo, pode ser melhor avaliado quando se realiza a pesquisa do HBV-DNA pela técnica de PCR

value of detection of HBVDNA in hepatocellular carcinoma patients using PCR technique

Thirty patients with primary HCC and 10 healthy controls were subjected to the study. Serological markers and HBV DNA were detected. In the patient group, HBsAg was positive in 21/30 [70 perecent] of cases,while anti-HBc was positive in 19/30 [63 percent] of cases. Both markers were positive in 17/30 [56,7 percent] of cases while both markers were negative in 7 of the cases. Only one patient of the control group was positive for HBsAg. Using the PCR technique to detect HBV. DNA in PBMC 27/30 [90 percent] of the patients group had detectable amounts, while none of the control group had HBV, DNA. This showed the close association between chronic HBV infection and disintegration of viral DNA which causes disarrangement of host DNA and produce malignant changes. The aim of this present work is to confirm the correlation between HBV and Hcc by detecting HBV-DNA by the polymerase chain reaction technique [PCR]