Detection and genotyping of human adenovirus and sapovirus in children with acute gastroenteritis in Belém, Pará, between 1990 and 1992: first detection of GI. 7 and GV. 2 sapoviruses in Brazil

Abstract INTRODUCTION: Acute gastroenteritis (AGE) is one of the most common causes of morbidity and mortality, especially among children from developing countries. Human adenovirus (HAdV) and sapovirus (SaV) are among the agents that cause AGE. The present study aimed to detect and genotype HAdV and SaV in 172 fecal samples from children with AGE, collected during a surveillance study carried out in a low-income community in Belém, Pará, between 1990 and 1992. METHODS: HAdV was detected by nested PCR, using primers Hex1deg/Hex2deg and NeHex3deg/NeHex4deg. SaV was assayed by reverse transcription PCR (RT-PCR), nested PCR, and quantitative PCR. The nucleotide sequence was determined by direct cycle sequencing. RESULTS: Overall, 43% (74/172) of samples were positive for HAdV, of which 70.3% (52/74) were sequenced and classified as belonging to five different species, mostly A and F. For SaV, positivity was 5.2% (9/172) and genotypes GI.1, GI.7, GII.1, and GV.2 were detected. CONCLUSIONS: The present results reinforce the need for further studies to obtain epidemiological data about the circulation of these viruses in Brazil, especially in the Amazon Region, where data from the early 1990's are scarce. Furthermore, the study describes for the first time the detection of SaV genotypes GI.7 and GV.2 in Brazil, showing that these types circulated in the region more than 25 years ago.

The role of human adenoviruses type 41 in acute diarrheal disease in Minas Gerais after rotavirus vaccination

Abstract Human adenovirus species F (HAdV-F) type 40 and 41 are commonly associated with acute diarrheal disease (ADD) across the world. Despite being the largest state in southeastern Brazil and having the second largest number of inhabitants, there is no information in the State of Minas Gerais regarding the role of HAdV-F in the etiology of ADD. This study was performed to determine the prevalence, to verify the epidemiological aspects of infection, and to characterize the strains of human adenoviruses (HAdV) detected. A total of 377 diarrheal fecal samples were obtained between January 2007 and August 2011 from inpatient and outpatient children of age ranging from 0 to 12 years. All samples were previously tested for rotavirus, norovirus, and astrovirus, and 314 of 377 were negative. The viral DNA was extracted, amplified using the polymerase chain reaction and the HAdV-positive samples were sequenced and phylogenetically analyzed. Statistical analyses were performed using the Chi-square test (p < 0.05), considering two conditions: the total of samples tested (377) and the total of negative samples for the remaining viruses tested (314). The overall prevalence of HAdV was 12.47% (47/377); and in 76.60% (36/47) of the positive samples, this virus was the only infectious agent detected. The phylogenetic analysis of partial sequences of 32 positive samples revealed that they all clustered with the HAdV-F type 41. The statistical analysis showed that there was no correlation between the onset of the HAdV infection and the origin of the samples (inpatients or outpatients) in the two conditions tested: the total of samples tested (p = 0.598) and the total of negative samples for the remaining viruses tested (p = 0.614). There was a significant association in the occurrence of infection in children aged 0–12 months for the condition 1 (p = 0.030) as well as condition 2 (p = 0.019). The occurrence of infections due to HAdV did not coincide with a pattern of seasonal distribution. These data indicate the significant involvement of HAdV-F type 41 in the etiology of ADD in Minas Gerais, which demonstrates the importance of other viral agents in the development of the disease after the introduction of rotavirus vaccine immunization.

Degradation and inactivation of adenovirus in water by photo-electro-oxidation / Degradação e inativação de adenovírus na água por fotoeletrooxidação

The present study analyzed the efficiency of the photo-electro-oxidation process as a method for degradation and inactivation of adenovirus in water. The experimental design employed a solution prepared from sterile water containing 5.107 genomic copies/L (gc/L) of a standard strain of human adenovirus type 5 (HAdV-5) divided into two equal parts, one to serve as control and one treated by photo-electro-oxidation (PEO) for 3 hours and with a 5A current. Samples collected throughout the exposure process were analyzed by real-time polymerase chain reaction (qPCR) for viral genome identification and quantitation. Prior to gene extraction, a parallel DNAse treatment step was carried out to assess the integrity of viral particles. Integrated cell culture (ICC) analyses assessed the viability of infection in a cell culture. The tested process proved effective for viral degradation, with a 7 log10 reduction in viral load after 60 minutes of treatment. The DNAse-treated samples exhibited complete reduction of viral load after a 75 minute exposure to the process, and ICC analyses showed completely non-viable viral particles at 30 minutes of treatment.

Resumo O presente estudo analisou a eficiência do processo de fotoeletrooxidação como metodologia para a degradação e inativação de adenovírus em água. A concepção experimental emprega uma solução preparada a partir de água estéril contendo 5,107 cópias genômicas/L (gc/L) de uma amostra padrão de adenovírus humano tipo 5 (HAdV-5), dividida em duas partes iguais, uma para servir como controle e outra tratada por fotoeletrooxidação (PEO) durante 3 horas e com uma corrente de 5A. As amostras recolhidas durante o processo de exposição foram analisadas por PCR quantitativo em tempo real (qPCR) para identificação e quantificação do genoma viral. Antes da extração de ácidos nucleicos, um passo de tratamento com DNAse paralelo foi realizado para avaliar a integridade das partículas virais. Um ensaio de qPCR integrado à cultura de células (ICC-qPCR) permitiu analisar a viabilidade de infecção em uma cultura de células. O processo mostrou-se eficaz testada para a degradação viral, com uma redução de 7 log10 da carga viral após 60 minutos de tratamento. As amostras tratadas com DNAse exibiram redução completa da carga viral após uma exposição de 75 minutos ao processo, e a análise de ICC-qPCR mostrou partículas virais completamente não-viáveis ​​em 30 minutos de tratamento.

Seasonal variation on the presence of adenoviruses in stools from non-diarrheic patients

Human adenoviruses (HAdV), members of the Adenoviridae family, are excreted through the fecal route and may be present in the feces of humans consuming contaminated food or water. The presence of HAdV from different serotypes in the feces of healthy individuals was already reported using conventional polymerase chain reaction; however, real-time PCR (qPCR) may reveal not only the rates of detection as well as demonstrate the viral loads excreted by healthy persons. Aiming to identify and characterize the presence of adenoviruses in stool samples, 147 fecal samples from patients with no records of diarrhea were analyzed (74 from winter season and 73 from summer) by Real-Time PCR (qPCR) assay and conventional PCR. HAdV genome was present in 43.8% (32/73) of stools samples collected during summer season and 21.6% (16/74) during winter. The rate of detection of genomic copies (gc) ranged from 4.04×10² to 6.72×10⁵gc/g of feces among the 147 samples analyzed, of which the ranged of genomic copies of DNA HAdV was major in summer. All samples were negative when tested for rotaviruses (RV) and noroviruses (NoV) by PCR conventional and qPCR respectively. HAdV is excreted constantly by infected individuals in the absence of clinical signs and the occurrence may vary seasonally.

Quantitative vs conventional PCR for detectionof human adenoviruses in water and sediment samples / PCR quantitativa versus convencional para a detecção de adenovírus humano em amostras de água e sedimento

SUMMARYHuman Adenoviruses (HAdV) are notably resistant in the environment. These agents may serve as effective indicators of fecal contamination, and may act as causative agents of a number of different diseases in human beings. Conventional polymerase chain reaction (PCR) and, more recently, quantitative PCR (qPCR) are widely used for detection of viral agents in environmental matrices. In the present study PCR and SYBR(r)Green qPCR assays were compared for detection of HAdV in water (55) and sediments (20) samples of spring and artesian wells, ponds and streams, collected from dairy farms. By the quantitative methodology HAdV were detected in 87.3% of the water samples and 80% of the sediments, while by the conventional PCR 47.3% and 35% were detected in water samples and sediments, respectively.

RESUMOOs adenovírus humanos (HAdV) são notavelmente resistentes ao ambiente. Estes agentes podem servir como indicadores efetivos de contaminação fecal, tanto quanto podem atuar como agentes causadores de diferentes doenças em seres humanos. A reação em cadeia da polimerase (PCR) e mais recentemente a PCR quantitativa (qPCR) são amplamente usadas para detecção de agentes virais em matrizes ambientais. No presente estudo, PCR e SYBR(r)Green qPCR foram comparadas para a detecção de HAdV em amostras de água (55) e sedimento (20) provenientes de nascentes, poços, açudes e arroios coletadas em propriedades leiteiras. A metodologia quantitativa detectou HAdV em 87,3% das amostras de água e 80% dos sedimentos, enquanto por PCR convencional a detecção foi de 47,3% e 35%, respectivamente.

Acute gastroenteritis and enteric viruses in hospitalised children in southern Brazil: aetiology, seasonality and clinical outcomes

Viral acute gastroenteritis (AG) is a significant cause of hospitalisation in children younger than five years. Group A rotavirus (RVA) is responsible for 30% of these cases. Following the introduction of RVA immunisation in Brazil in 2006, a decreased circulation of this virus has been observed. However, AG remains an important cause of hospitalisation of paediatric patients and only limited data are available regarding the role of other enteric viruses in these cases. We conducted a prospective study of paediatric patients hospitalised for AG. Stool samples were collected to investigate human adenovirus (HAdV), RVA, norovirus (NoV) and astrovirus (AstV). NoV typing was performed by nucleotide sequencing and phylogenetic analysis. From the 225 samples tested, 60 (26%) were positive for at least one viral agent. HAdV, NoV, RVA and AstV were detected in 16%, 8%, 6% and 0% of the samples, respectively. Mixed infections were found in nine patients: HAdV/RVA (5), HAdV/NoV (3) and HAdV/NoV/RVA (1). The frequency of fever and lymphocytosis was significantly higher in virus-infected patients. Phylogenetic analysis of NoV indicated that all of these viruses belonged to genotype GII.4. The significant frequency of these pathogens in patients with AG highlights the need to routinely implement laboratory investigations.

Detection of human adenovirus, rotavirus and enterovirus in water samples collected on dairy farms from Tenente Portela, Northwest of Rio Grande do Sul, Brazil

Viral gastroenteritis and other waterborne diseases are a major concern for health in Brazil. A number of studies were conducted about the presence of viruses on water samples from Brazilian areas. However, the knowledge about the occurrence of viral contamination of drinking water sources in rural settings of the country is insufficient. On the present work, 15 samples from 5 dairy farms located at the municipality of Tenente Portela were collected and analysed for the presence of human adenoviruses (HAdV), as well as human enteroviruses (EV) and rotaviruses (RV). HAdV was present on 66.66% of the water samples, and have been found in all samples from artesian wells and springs, which are used as sources of drinking water for the individuals inhabiting those farms. EV and RV found only in one sample each. The detection rates of HAdV on the water from these dairy farms are alarming and point towards a situation of elevated environmental contamination by fecal microorganisms of human origin and poor basic sanitation conditions.

Human adenovirus detection among immunocompetent and immunocompromised patients presenting acute respiratory infection

INTRODUCTION: Human adenoviruses (HAdV) play an important role in the etiology of severe acute lower respiratory infection, especially in immunocompromised individuals. The aim of the present study was detect the HAdV through different methods: direct fluorescence assay (DFA) and nested-polymerase chain reaction (PCR-nested) from patients with acute respiratory infection (ARI) up to 7 days of symptoms onset. METHODS: Samples (n=643) were collected from different risk groups during from 2001 to 2010: 139 adults attended in an Emergency Room Patients (ERP); 205 health care workers (HCW); 69 from Renal Transplant Outpatients (RTO); 230 patients in hematopoietic stem cell transplantation (HSCT) program. RESULTS: Among all patients (n=643) adenovirus was detected on 13.2% by DFA and/or PCR: 6/139 (4.3%) adults from ERP, 7/205 (3.4%) from HCW samples, 4/69 (5.8%) from RTO and 68/230 (29.5%) from HSCT patients. Nested PCR showed higher detection (10%) compared to DFA test (3.8%) (p < 0.001). HSCT patients presented significantly higher prevalence of HAdV infection. CONCLUSIONS: Adenovirus detection through nested-PCR assay was higher. However the inclusion of molecular method in laboratorial routine diagnostic should be evaluated considering the reality of each specific health service. .

Isolation of a variant human adenovirus identified based on phylogenetic analysis during an outbreak of acute keratoconjunctivitis in Chennai.

Background & objectives: Though several viruses are responsible for conjunctivitis, but human adenovirus (HAdV) is by far the most common cause. Epidemic conjunctivitis causes morbidity and early detection of aetiological agent is essential in preventing spread of disease as some of serotypes of adenoviruses cause a severe form of conjunctivitis. This study was undertaken to identify the causative agent of conjunctivitis outbreak in Chennai in 2010. Methods: Conjunctival samples collected from 17 patients with conjunctivitis were subjected to virological investigations. Culture and PCR for detection of adenovirus and enterovirus were carried out. PCR positive products were further subjected for DNA sequencing. The nucleotide sequences of the hexons of isolates were analyzed by comparison with all 51 human adenovirus strains. Phylogenetic tree was constructed using DAMBE software. Results: Among 17 patients, seven were positive for adenovirus by PCR on the direct specimen, none was positive for enterovirus. Eleven of 30 conjunctival swabs showed cytopathic effect in HEp-2 cell line and were confirmed as HAdV by PCR. The DNA sequence data of the 11 isolates had equal percentage of homology with HAdV 6 and 2 on blast analysis. On phylogenetic analysis with GeneBank data of 51 adenovirus strains, 11 isolates from patients during the outbreak of conjunctivitis formed a separate clade indicating a new variant strain. Interpretation & conclusions: Based on phylogenetic analysis it was concluded that the recent conjunctivitis outbreak that occurred in Chennai was caused by a variant adenovirus strain.

Gravidade das coinfecções virais em lactentes hospitalizados com infecção por vírus sincicial respiratório / Severity of viral coinfection in hospitalized infants with respiratory syncytial virus infection

OBJETIVO: Comparar a gravidade de infecções causadas por um único vírus (VSR) com a gravidade de coinfecções. MÉTODOS: Este estudo avaliou uma coorte histórica de lactentes com infecção aguda por VSR. Secreção de nasofaringe foi coletada de todos os pacientes rotineiramente para pesquisa viral usando técnicas de biologia molecular. Os seguintes desfechos foram analisados: tempo total de internação, duração da oxigenioterapia, admissão em unidade de terapia intensiva e uso de ventilação mecânica. Os resultados foram ajustados para os fatores confundidores (prematuridade, idade e aleitamento materno). RESULTADOS: Foram incluídos no estudo 176 lactentes com idade média de 4,5 meses e diagnósticos de bronquiolite e/ou pneumonia. Cento e vinte e um tinham infecção única por VSR, e 55 tinham coinfecções (24 VSR + adenovírus, 16 VSR + metapneumovírus humano e 15 outras associações menos frequentes). Os quatro desfechos de gravidade avaliados foram semelhantes entre o grupo com infecção única por VSR e os grupos com coinfecções, independente do tipo de vírus associado com o VSR. CONCLUSÃO: As coinfecções virais não parecem alterar o prognóstico de lactentes hospitalizados com infecção aguda por VSR.

OBJECTIVE: To compare the severity of single respiratory syncytial virus (RSV) infections with that of coinfections. METHODS: A historical cohort was studied, including hospitalized infants with acute RSV infection. Nasopharyngeal aspirate samples were collected from all patients to detect eight respiratory viruses using molecular biology techniques. The following outcomes were analyzed: duration of hospitalization and of oxygen therapy, intensive care unit admission and need of mechanical ventilation. Results were adjusted for confounding factors (prematurity, age and breastfeeding). RESULTS: A hundred and seventy six infants with bronchiolitis and/or pneumonia were included in the study. Their median age was 4.5 months. A hundred and twenty one had single RSV infection and 55 had coinfections (24 RSV + adenovirus, 16 RSV + human metapneumovirus and 15 other less frequent viral associations). The four severity outcomes under study were similar in the group with single RSV infection and in the coinfection groups, independently of what virus was associated with RSV. CONCLUSION: Virus coinfections do not seem to affect the prognosis of hospitalized infants with acute RSV infection.

Adenovirus respiratory infection: significant increase in diagnosis using PCR comparing with antigen detection and culture methods / Infecção respiratória aguda por adenovirus: comparação dos métodos de PCR e imunofluorescência indireta para o seu diagnóstico e dados clínicos dos pacientes infectados

Adenovirus (AdV) respiratory infections are usually described as being associated with high mortality rates. Laboratory diagnosis is essential for the establishment of the appropriate therapy, and for guiding the implementation of preventive measures in order to prevent the spread of the infection. Aiming to analyze the sensitivity and specificity of the laboratorial diagnosis methods available, we compared antigen detection by indirect immunofluorescence assay (IF), and a specific nested polymerase chain reaction (PCR), to detect AdV in respiratory samples collected from patients admitted to hospital with acute respiratory disease. Positive samples were inoculated into a cell culture to confirm the results. We analyzed 381 samples from the nasopharyngeal aspirates collected during the year 2008; of these, 2.6 percent tested were positive for adenovirus through IF and 10 percent through PCR; positive isolation was obtained in 40 percent and 26 percent of these cases, respectively. Most infected patients were children under six months of age, and despite of the fact that a significant number of patients required intensive care, the mortality rate was low (5 percent). In conclusion, molecular methods were found to be useful for rapid diagnosis of adenovirus infections with higher sensitivity than antigen detection; their introduction permitted a significant increase in diagnoses of adenovirus infections.

Infecções respiratórias por Adenovírus (ADV) são geralmente descritas associadas com alta mortalidade. O diagnóstico laboratorial é essencial para o estabelecimento da terapêutica adequada e para orientar a implantação de medidas preventivas evitando a propagação da infecção. Com o objetivo de analisar a sensibilidade e a especificidade dos métodos de avaliação de diagnóstico laboratorial, foi comparada a detecção de antígeno por imunofluorescência indireta (IF) com a reação em cadeia da polimerase específica (PCR) para detectar AdV em amostras respiratórias coletadas de pacientes internados com doença respiratória aguda. As amostras com resultados positivos foram inoculadas em cultura celular. Foram analisadas 381 amostras da secreção nasofaríngea coletadas durante o ano de 2008, das quais 2,6 por cento foram positivas pela IF e 10 por cento pela PCR, isolamento positivo foi obtido em 40 por cento e 26 por cento dos casos positivos pelos testes anteriores, respectivamente. A maioria dos pacientes infectados eram crianças com menos de seis meses de idade, e apesar do fato de que um número significativo de pacientes necessitou de cuidados intensivos, a taxa de mortalidade foi baixa (5 por cento). Em conclusão, os métodos moleculares são úteis para o diagnóstico rápido de infecções por adenovírus com maior sensibilidade do que a detecção do antígeno, a sua introdução na rotina permitiu um aumento significativo no diagnóstico de infecções por adenovírus.

Queratoconjuntivitis por adenovirus generadas a partir de una consulta oftalmológica / Adenovirus keratoconjunctivitis acquired in an ophtalmology clinic

Introduction: Eye infection is a common cause of ophtalmologic consultation. Adenovirus keratoconjunctivitis outbreaks are common worldwide but its impact and clinical characteristic in Chilean population is unkown. Objective: To describe a series of adenovirus keratoconjunctivitis cases. Patients and Method: The Índex case and contacts received medical care in the Hospital Clínico Universidad de Chile between April and August 2006. A complete ophthalmologic exam and microbiologic evaluation was performed. Results: Nine patients presented a pattern of characteristic epidemic keratoconjunctivitis. In x cases sub-corneal epithelial infiltrates were observed for a period of more than six months. Three affected patients were ophtalmologists, staff at the Hospital. In seven patients ADV was isolated all bellonging to type D genus. Conclusions: Adenovirus type D caused epidemic keratoconjunctivitis in a series of Chilean individuals. Ophthalmologist may have transmitted the virus to patients.

Introducción: La patología ocular infecciosa es frecuente en la consulta oftalmológica, especialmente la conjuntivitis y queratoconjuntivitis epidémica (QCE). Brotes de esta patología son causados por adenovirus (ADV) en el extranjero; en Chile se desconoce su impacto y características. Objetivos: Describir una serie de casos de queratonconjuntivitis epidémica por adenovirus. Material y Pacientes: Al caso índice y los contactos de una serie de casos de QCE por ADV que consultaron en el Hospital Clínico de la Universidad de Chile, entre abril y agosto de 2006, se les realizó examen oftalmológico completo y estudio de ADV por aislamiento viral, detección de antígenos y de genoma viral. Se estableció el género de ADV mediante reacción de polimerasa en cadena. Resultados: Los 9 pacientes infectados presentaron QCE característica. En algunos casos se observaron infiltrados sub-epiteliales corneales que se extendieron por más de seis meses. Tres pacientes eran médicos oftalmólogos. En 7 de los 9 pacientes examinados se aisló ADV; todos del género D. Conclusiones: En Chile, la QCE puede ser causada por el subgénero tipo D. El médico oftalmólogo es un potencial vector en la transmisión de ADV en un brote de QCE, por lo que es fundamental que sea considerado en las estrategias de prevención de esta patología.

Evaluation of HA negatively charged membranes in the recovery of human adenoviruses and hepatitis A virus in different water matrices

Human adenoviruses (HAdV) and hepatitis A virus (HAV) are shed in the faeces and consequently may be present in environmental waters, resulting in an increase in pathogen concentration that can affect water quality and human health. The aim of this study was to evaluate an adsorption-elution method which utilizes negatively charged membrane HA to determine the efficient recovery of HAdV and HAV from different water matrices and to combine this procedure with a qualitative molecular method (nested RT-PCR and nested PCR). The best efficiency recovery was achieved in distilled water and treated wastewater effluent (100 percent) for both viruses and in recreational lagoon water for HAV (100 percent). The efficiency recovery was 10 percent for HAdV and HAV in seawater and 10 percent for HAdV in lagoon water. The viral detection limit by nested PCR for HAV in water samples ranged between 20-0.2 FFU/mL and 250 and 25 TCID50/mL for HAdV. In conclusion, these results suggest that the HA negatively charged membranes vary their efficiency for recovery of viral concentration depending upon the types of both enteric viruses and water matrices.

Comparative study of two extraction methods for enteric virus recovery from sewage sludge by molecular methods

The aim of this study was to compare two nucleic acid extraction methods for the recovery of enteric viruses from activated sludge. Test samples were inoculated with human adenovirus (AdV), hepatitis A virus (HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an adsorption-elution-precipitation method. Two extraction methods were used: an organic solvent-based method and a silica method. The organic-based method was able to recoup 20 percent of the AdV, 90 percent of the RV and 100 percent of both the PV and HAV from seeded samples. The silica method was able to recoup 1.8 percent of the AdV and 90 percent of the RV. These results indicate that the organic-based method is more suitable for detecting viruses in sewage sludge.

Longitudinal study on occurrence of adenoviruses and hepatitis A virus in raw domestic sewage in the city of Limeira, São Paulo / Estudo longitudinal da ocorrência de adenovírus e vírus da hepatite A em esgoto doméstico na cidade de Limeira, SP

The aim of this study was to verify the presence and annual distribution of adenoviruses and hepatitis A virus in domestic sewage in the city of Limeira, São Paulo. Fifty samples with a volume of 8 liters each were collected weekly from December 2004 to December 2005. The viruses were concentrated by filtration through positively charged ZP60S filter membranes, followed by ultracentrifugation. Human adenoviruses (HAdV) were detected by PCR followed by nested-PCR and screening for species F was done by restriction of the PCR product with TaqI endonuclease. Virus infectivity assays were performed by inoculation of concentrates onto HEp-2 cell monolayers. RT-PCR was used for the detection of hepatitis A virus. HAdV were detected in all samples, and 64% of samples were positive for infectious virus. Species F was present in 82% of the samples. Hepatitis A virus was detected in 48% of the samples. These results demonstrate that HAdV and HAV were present in the domestic sewage of Limeira throughout the period of study, demonstrating the importance of an adequate treatment before the disposal in the environment.

O objetivo do estudo foi verificar a ocorrência e a distribuição anual de adenovírus humanos e vírus da Hepatite A (VHA) no efluente doméstico da cidade de Limeira, São Paulo, ao longo do período de Dezembro de 2004 e Dezembro de 2005, com vistas à futura implementação de sistemas de tratmento de água de esgoto. Cinquenta amostras de efluente bruto com volume de 8L cada foram colhidas semanalmente e os vírus concentrados por filtração em membrana eletropositiva ZP60S, seguida de ultracentrifugação. Adenovírus foram detectados por PCR e nested-PCR. Adenovírus da espécie F foram distinguidos das demais por restrição do produto da PCR com endonuclease TaqI. Ensaios de infectividade viral foram realizados em culturas de células HEp-2. A presença do vírus da hepatite A também foi pesquisada nas mesmas amostras, fazendo-se uso de método de RT-PCR. Adenovírus foram detectados em todas as amostras, sendo a espécie F identificada em 82% destas. Sessenta e quatro por cento dos adenovírus detectados ainda estavam infecciosos. O vírus da Hepatite A foi detectado em 48% das amostras examinadas. Estes resultados evidenciam a presença e a circulação de Adenovírus humano e VHA nas águas de esgoto doméstico de Limeira ao longo do período de estudo, demonstrando a importância de um tratamento adequado desse material antes da disposição no meio ambiente.

Aislamiento y caracterización de cepas de Adenovirus que colonizan el tracto gastrointestinal de pacientes cubanos seropositivos al VIH / Isolation and characterization of Adenovirus strains that invade the gastrointestinal tract of Cuban HIV-positive patients

Introducción: el tracto gastrointestinal es uno de los sistemas que se afecta con frecuencia en el paciente seropositivo al virus de la inmunodeficiencia humana (VIH), de manera que las alteraciones o enfermedades en este sitio representan una causa importante de morbilidad y mortalidad. El hallazgo de cepas de adenovirus, raramente aisladas en pacientes inmunocompetentes, ha sido asociado en ocasiones a la presencia de diarrea, lo cual refleja la importancia del estudio y la tipificación de estas cepas. Objetivos: conocer la prevalencia de los Adenovirus en muestras de heces fecales de pacientes infectados con el VIH, con diarrea aguda o crónica, así como la caracterización de los agentes aislados, mediante la implementación de métodos rápidos y confiables. Métodos: fueron analizadas 167 muestras de heces fecales de individuos seropositivos al VIH y 127 seronegativos como grupo control. La prevalencia fue investigada mediante el aislamiento en cultivo celular, se realizó además la tipificación de las cepas aisladas, mediante técnicas de biología molecular como la reacción en cadena de la polimerasa y la secuenciación nucleotídica. El análisis estadístico se efectuó mediante la prueba exacta de Fisher contenido en el paquete estadístico Epi Info (Versión 6.04). Resultados: la prevalencia en los pacientes VIH+ fue de 15 por ciento, mientras que en los VIH- fue de 4 por ciento; la diferencia resultó estadísticamente significativa; 96 por ciento de las cepas aisladas en los pacientes VIH+ correspondieron al subgénero D, asociadas en 62,5 por ciento a cuadros diarreicos y estadios avanzados de la infección por VIH. Conclusiones: los resultados sugieren la importancia de incluir a los adenovirus y su tipificación en el diagnóstico de rutina de los pacientes VIH+, lo cual sería de interés en el manejo clínico de la infección y aportaría datos claves en la vigilancia y el control de la transmisión de la infección hospitalaria y los análisis epidemiológicos.

Bacckground: the gastrointestinal tract is one of the most frequently affected systems in HIV-positive patients, therefore, changes or diseases occurring in this site represent an important cause of morbidity and mortality. The finding of adenovirus strains, rarely isolated in immunocompetent patients, has been occasionally associated to diarrheas, which reflects the importance of study and typing of these strains. Objectives: to find out the prevalence of Adenovirus in feces samples from HIV patients having chronic or acute diarrheas, as well as the characterization of isolated agents through the application of reliable and quick methods. Methods: one hundred and sixty seven fecal samples from HIV-positive patients and 127 from seronegative subjects as the control group were analyzed. Isolation in cell cultures determined the prevalence and molecular biology techniques as the polymerase chain reaction and nucleotide sequencing allowed typing of isolated strains. The statistical analysis was based on the Fisher´s exact test included in Epi Info (6.04) statistical software. RESULTS: the prevalence in HIV positive patients was 15 percent whereas in the HIV-negative was 4 percent. The difference was statistically significant; 96 percent of isolated strains in HIV positive patients belonged to subgenus D, which are linked in 62.5 percent of cases to acute diarrheal condition and advanced stages of HIV infection. Conclusions: the results indicated the importance of including adenovirus and their typing in the routine diagnosis of patients positive to HIV, all of which would be useful in the clinical management of infection and would contribute key data on surveillance and control of hospital infection transmission and the epidemiological analysis.

Adenovirus, calicivirus and astrovirus detection in fecal samples of hospitalized children with acute gastroenteritis from Campo Grande, MS, Brazil

We analyzed fecal samples from hospitalized children up to three years of age with acute gastroenteritis at Campo Grande, Mato Grosso do Sul, Brazil, from May 2000-January 2004. Astrovirus and calicivirus were detected by Reverse Transcription-Polymerase Chain Reaction and adenovirus was detected using the Rotavirus and Adenovirus combined immunoenzyme assay. Astrovirus, adenovirus and calicivirus were detected at rates of 3.1 percent, 3.6 percent and 7.6 percent, respectively. These results re-emphasize the need for the establishment of regional vigilance systems to evaluate the impact of enteric viruses on viral gastroenteritis.

Adenoviruses C in non-hospitalized Mexican children older than five years of age with acute respiratory infection

Adenoviruses (AdV) are commonly involved in acute respiratory infections (ARI), which cause high morbidity and mortality in children. AdV are grouped in six species (A-F), which are associated with a wide range of diseases. The aim of this study was to identify the AdV species infecting non-hospitalized Mexican children with ARI symptoms, attending to the same school. For that, a PCR/RFLP assay was designed for a region of the hexon gene, which was chosen, based on the bioinformatical analysis of AdV genomes obtained from GenBank. A total of 100 children's nasopharyngeal samples were collected from January to June, 2005, and used for viral isolation in A549 cells and PCR/RFLP analysis. Only 15 samples produced cytopathic effect, and in all of them AdV C was identified. AdV C was also identified in eight additional nasopharyngeal samples which were negative for viral isolation. In summary, this outpatient population showed a rate of AdV infection of 23 percent, and only AdV C was detected.

Antigenic and genomic characterization of adenovirus associated to respiratory infections in children living in Northeast Brazil

From January to December 1998, nasopharyngeal aspirates were obtained from 482 children with acute respiratory infections attended in emergence department and wards of a teaching hospital in the city of Salvador, Brazil. The samples were tested for the presence of adenovirus by isolation in tissue culture and indirect immunofluorescence assay. Eleven adenoviruses were detected by both methods in the same clinical samples. Infections by adenovirus were observed during seven months of the year without association with rainy season. Genome analysis was performed on these 11 isolates. Species C was represented by serotypes 1, 2 and 5. Within species B, only serotype 7 (Ad7) was detected. Two genomic variants of Ad1, two variants of Ad2, one of Ad5, and one of Ad7 (7h) were identified. This is the first study of molecular epidemiology of adenovirus associated to acute respiratory infections in children living in Northeast Brazil, and contributes to a better understanding of adenovirus infections in the country.

Diagnóstico de conjuntivite adenoviral pelo RPS Adenodetector® / Adenovirus conjunctivitis diagnosis using RPS Adenodetector®

OBJETIVO: Avaliar a utilização do RPS Adenodetector®, como método diagnóstico de pacientes com quadro clínico de conjuntivite adenoviral. MÉTODOS: Análise de série de casos consecutivos de pacientes com diagnóstico clínico de ceratoconjuntivite adenoviral submetidos comparativamente ao teste RPS Adenodetector® e a raspado conjuntival para cultura de vírus. RESULTADOS: Dos 11 pacientes avaliados, 10 pacientes apresentavam acometimento unilateral. Em relação ao tempo de início dos sintomas no momento da colheita, 5 (45,5 por cento) pacientes apresentavam dois dias de história, 5 (45,5 por cento) apresentavam três dias e 1 (9,1 por cento) apresentava 7 dias. A cultura para adenovírus foi positiva em 8 pacientes (73 por cento) e o RPS Adenodetector® foi positivo em 9 pacientes (82 por cento). Oito pacientes apresentaram o teste rápido e cultura positiva. Um paciente apresentou teste RPS Adenodetector® positivo com cultura negativa. Os dois pacientes com teste RPS Adenodetector® negativo apresentaram cultura negativa. O RPS Adenodetector® mostrou sensibilidade de 100 por cento e especificidade de 67 por cento adotando-se a cultura de vírus como exame padrão-ouro para o diagnóstico de conjuntivite adenoviral. CONCLUSÃO: O RPS Adenodetector® foi útil para o diagnóstico de conjuntivite adenoviral e pode auxiliar na orientação do paciente quanto ao contágio e disseminação da doença.

PURPOSE: To evaluate the RPS Adenodetector®, a rapid immunochromatographic test, in the diagnosis of patients with clinical overt adenoviral conjunctivitis. METHODS: Consecutive case series. Patients underwent conjunctiva scraping for RPS Adenodetector® test and culture to identify adenovirus. RESULTS: A total of 11 patients were studied, and 10 had unilateral disease. Five (45.5 percent) had symptoms for 2 days, 5 for three days, and 1 for 7 days. Adenovirus culture was positive in 8 patients (73 percent) and RPS Adenodetector® was positive in 9 (82 percent) patients. Eight patients had adenovirus identification by both methods. In one patient the RPS Adenodetector® was positive in contrast to a negative culture. The two patients revealing negative RPS Adenodetector® results also had negative cultures. The sensitivity was 100 percent and the specificity was 67 percent. CONCLUSION: The RPS Adenodetector® is a useful tool in the rapid diagnosis of adenovirus conjunctivitis and may contribute to the spread control of this highly contagious disease.