Analysis of enterovirus infection type among acute respiratory tract infection cases in Luohe City, Henan Province from 2017 to 2021 / 中华预防医学杂志

Objective: To understand the infection status of Enterovirus (EV) in cases of acute respiratory infections (ARIs) in Luohe City, Henan Province from 2017 to 2021, and analyze the prevalence and type composition of EV in ARIs. Methods: From October 2017 to May 2021, pharyngeal swab samples were collected from 1 828 patients with ARIs in Luohe Central Hospital and the clinical epidemiological data of these cases were also collected. EV-positive samples were identified by Quantitative Real-time Polymerase Chain Reaction (qPCR). The 5'-untranslated region (5'UTR) was amplified by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The results of 5'UTR region were initially typed by Enterovirus Genotyping Tool Version 1.0. Based on the typing results, the full-length of VP1 region was amplified by RT-PCR. The EV typing was identified again by VP1 region. Results: Among 1 828 cases of ARIs, 56.7% (1 036) were males. The median (Q1, Q3) age was about 3 (1, 5) years. Patients under 5 years old accounted for 71.6% (1 309 cases). Among all cases, a total of 71 EV-positive samples were identified by qPCR, with a detection rate of 3.88% (71/1 828). The EV detection rates for men and women were 3.28% (34/1 036) and 4.67% (37/792), without statistically significant differences (χ2=2.32, P=0.14). The EV detection rates for 2 to <6 years, 6 months to <2 years, 6 to <10 years, and <6 months were 6.29% (48/763), 3.00% (18/600), 2.52% (4/159), and 1.67% (1/60) (χ2=27.91, P<0.001). The EV detection rate was 0.92% (3/326) in autumn and winter of 2017. The EV detection rates were 1.18% (6/508), 2.47% (12/485) and 8.31% (34/409) in each year from 2018 to 2020, with an increasing trend year by year(χ2trend=29.76, P<0.001). The main prevalent seasons were summer and autumn. The detection rate in spring of 2021 was 4.00% (4/100). A total of 12 types were identified and classified as CVA2, CVA4, CVA5, CVA6, CVA10, CVB3, CVB5, E5, E11, E30, PV-1, and EV-D68. The types of CVA2, CVA10, CVA6, and CVB3 were the dominant phenotypes. In 59 sample of EV typing, the main clinical manifestation was upper respiratory tract infection (36/59, 61.01%). The dominant types detected in upper respiratory tract infections were CVA10 (10/36, 27.78%), CVA6 (9/36, 25.00%) and CVB3 (8/36, 22.22%). The dominant type detected in lower respiratory tract infections was CVA2 (7/19, 36.84%). Conclusion: In Luohe City, Henan Province from 2017 to 2021, EV infection in ARIs cases has clear seasonal and age-specific patterns, and the dominant types of upper and lower respiratory tract infections are different.

Arbovirus, herpesvirus, and enterovirus associated with neurological syndromes in adult patients of a university hospital, 2017-2018

Abstract INTRODUCTION: Herpesviruses, enteroviruses, and arboviruses are important because of their clinical relevance and ability to cause meningitis, encephalitis, meningoencephalitis, and other diseases. The clinical virology associated with diagnostic technologies can reduce the morbidity and mortality of such neurological manifestations. Here we aimed to identify the genomes of agents that cause neurological syndromes in cerebrospinal fluid (CSF) samples from patients with suspected nervous system infections admitted to the University Hospital of the University of Campinas, São Paulo, Brazil, in 2017-2018. METHODS: CSF samples collected from adult patients with neurological syndrome symptoms and negative CSF culture results were analyzed using polymerase chain reaction (PCR), reverse transcriptase-PCR, and real-time PCR, and their results were compared with their clinical symptoms. One CSF sample was obtained from each patient. RESULTS: Viral genomes were detected in 148/420 (35.2%) CSF samples: one of 148 (0.2%) was positive for herpes simplex virus-1; two (0.5%) for herpes simplex virus-2; eight (1.9%) for varicella-zoster virus; four (1%) for Epstein-Barr virus; one (0.2%) for cytomegalovirus; 32 (7.6%) for human herpesvirus-6; 30 (7.1%) for non-polio enterovirus; 67 (16.0%) for dengue virus, three (0.7%) for yellow fever virus, and 21 (5%) for Zika virus. CONCLUSIONS: The viral genomes were found in 35.2% of all analyzed samples, showing the high prevalence of viruses in the nervous system and the importance of using a nucleic acid amplification test to detect viral agents in CSF samples.

Enterovirus asociados a Afecciones del Sistema Nervioso Central confirmados por biología molecular / Enteroviruses associated with Central Nervous System Infections confirmed by molecular biology

Aproximadamente 90% de las meningitis asépticas son causadas por enterovirus (EV), miembro de la familia de los picornavirus. Los EV son ubicuos, se diseminan por vía fecal-oral y contacto directo, responsables de brotes o casos esporádicos con importante morbilidad. El diagnóstico se basa en la presentación clínica, imágenes, estudio citoquímico del líquido cefalorraquídeo (LCR) y la Reacción en Cadena de la Polimerasa (PCR), gold estándard que reemplaza al aislamiento viral y la serología. El objetivo de este estudio descriptivo de corte transversal fue determinar la presencia de EV por RT-PCR en el LCR de pacientes con sospecha clínica de meningitis aséptica, internados en servicios públicos y privados de Asunción y departamento Central del Paraguay de noviembre de 2007 a noviembre de 2014. El material genético fue extraído mediante el mini kit ADN y ARN Qiagen® que fue sometido a RT-PCR. Se incluyeron LCR de 203 pacientes, 124 (61%) niños (4 días-15 años) y 79 (39%) adultos (16-81 años). Setenta y siete (38%) provenían de servicios públicos y 126 (62%) de privados; 115 (57%) fueron varones. Se detectó RNA de EV en 166 (82%) pacientes, 90 niños y 76 adultos, y mayor número de casos entre los meses de octubre a abril. Este es el primer trabajo en el país y muestra una importante participación del EV en pacientes por infecciones del SNC compatibles con meningitis asépticas de etiología viral. La sospecha clínica fue mayor en niños, sin embargo la proporción de resultados positivos fue mayor en adultos. Se observó mayor circulación en los meses cálidos(AU)

Identification of enteric viruses circulating in a dog population with low vaccine coverage

ABSTRACT Although the use of vaccines has controlled enteric diseases in dogs in many developed countries, vaccine coverage is still under optimal situation in Brazil. There is a large population of nonimmunized dogs and few studies about the identification of the viruses associated with diarrhea. To address this situation, stool samples from 325 dogs were analyzed by polymerase chain reaction for the detection of common enteric viruses such as Canine adenovirus (CAdV), Canine coronavirus (CCoV), Canine distemper virus (CDV), Canine rotavirus (CRV) and Carnivorous protoparvovirus 1 (canine parvovirus 2; CPV-2). At least one of these species was detected in 56.6% (184/325) of the samples. The viruses detected most frequently in either diarrheic or nondiarrheic dog feces were CPV-2 (54.3% of the positive samples), CDV (45.1%) and CCoV (30.4%), followed by CRV (8.2%) and CAdV (4.9%). Only one agent was detected in the majority of the positive samples (63%), but co-infections were present in 37% of the positive samples and mainly included CDV and CPV-2. The data presented herein can improve the clinical knowledge in regions with low vaccine coverage and highlight the need to improve the methods used to control these infectious diseases in domestic dogs.

Molecular identification of viral agents associated with acute conjunctivitis: a prospective controlled study

Abstract Background: Viral conjunctivitis are the most frequent infections in ophthalmology clinics. The diagnosis is usually relying on clinical findings and medical history. However, topical antibiotics are often used unnecessarily addition to symptomatic treatment because of unsure agents. We aimed to detect the Adenovirus, Coxsackievirus and Enterovirus from conjunctiva and pharyngeal samples of patients. Methods: The conjunctiva and pharyngeal samples of the patients with conjunctivitis were taken by Virocult transport media and kept at -80 ºC up to study day. Adenovirus spp, Enterovirus 70 and Enterovirus 71, Coxsackie A24 and Coxsackie A16 were detected by real-time PCR. Samples from healthy health care workers of ophthalmology clinic were used for control group. Results: A total of 176 samples (conjunctival and pharyngeal samples of 62 patient and 26 healthy subjects) were included. The mean age of 34 (55.7%) male and 27 (44.3%) female patients was 34 ± 17. Twenty five (40.3%) of the patients were receiving antibiotic drops at first visit. The main etiologic agent in conjunctival samples was found to be Adenovirus (46/62, 74.2%) followed by Enterovirus 70 (4/62, 6.4%) and Enterovirus 71 (4/62, 6.4%). Coxsackievirus 16 and 24 were also found in 2 patients (1/62 each, 1.6%). Pharyngeal samples were also positive for Adenovirus (20/62, 32.3%), Enterovirus 70 and 71 (7/62, 11.3% and 5/62, 8.1% respectively), Coxsackievirus 16 and 24 (2/62, 3.2% and 1/61, 1.6%). Conclusions: It is very difficult in viral conjunctivitis to make clinical differentiation caused by different agents because of common clinical signs and symptoms. In routine clinical work, the viral conjunctivitis usually related with Adenovirus. But almost one fourth of the patients' conjunctivitis were not related to Adenovirus, which shows the importance of the laboratory diagnostics. True diagnosis plays an important role on prevention of contamination and unnecessary use of antibiotics in viral conjunctivitis.

Moving beyond classical markers of water quality: detection of enteric viruses and genotoxicity in water of the Sinos River / Indo além dos marcadores clássicos de qualidade da água: detecção de vírus entéricos e genotoxicidade na água do Rio dos Sinos

It is well recognized that the classical biological and chemical markers of environmental pollution do not necessarily indicate the presence or absence of emerging threats to public health, such as waterborne viruses and genotoxicants. The purpose of this preliminary study was to evaluate the presence of material of enteroviruses (EV), rotavirus (RV) and adenovirus (AdV) and genotoxicity in water samples from points of routine monitoring of water quality in the main course of the Sinos River. The points are classified into different levels of pollution in accordance to the Brazilian federal regulations. Viral genomes from EV, AdV were detected in two of the 4 collection points regardless of the level of urbanisation of the surrounding areas. In contrast, genotoxicity was not observed in piava (Leporinus obtusidens) fingerlings cultivated on these same water samples. Results were compared with classical physical, chemical and microbiological parameters. There was no clear evidence of association between any of the classical markers and the presence of viral genomes in the water samples tested.

É amplamente reconhecido que os marcadores biológicos e químicos clássicos para a poluição ambiental não necessariamente indicam a presença ou ausência de ameaças emergentes à saúde pública, tais como vírus transmitidos pela água e genotoxicantes. Este estudo preliminar teve por objetivo detectar material genético de enterovírus (EV), rotavírus (RV) e adenovírus (AdV) e genotoxicidade em amostras de água de pontos de monitoramento de rotina da qualidade da água no curso principal do rio dos Sinos. Os pontos são classificados em níveis diferentes de poluição, de acordo com as normativas federais brasileiras. Genomas virais de EV, RV e RV foram detectados em dois dos quatro pontos de coleta, independente do nível de urbanização das áreas adjacentes. Por outro lado, não foi observada genotoxicidade em alevinos de piava (Leporinus obtusidens) cultivados nestas mesmas amostras de água. Os resultados são comparados com marcadores físicos, químicos e microbiológicos clássicos, não há nenhuma evidência clara da associação entre qualquer um dos marcadores clássicos e da presença de genomas virais nas amostras de água testadas.

La vacuna polio oral en lactantes no interfiere con la detección de enterovirus en sangre / Oral polio vaccine in infants does not interfere in detection of enterovirus in blood

Introduction: There is not known if a viraemia post-oral polio vaccine (OPV) is detectable by modern molecular techniques. Such viraemia could affect the performance of the real time-polymerase chain reaction (PCR) for non polio enterovirus (EV) detection, technique of growing clinical use for the study of febrile infants. Objective: To determine viraemia post-first dose of OPV in healthy infants, by molecular techniques. Patients and Methods: 50 infants less than three months without previous VPO were randomized in 5 groups: a control group with pre-vaccination blood sample (BS), group 1 BS at day 2, group 2 BS at day 4, group 3, BS at day 6 and group 4, BS at day 8 post-vaccination. Conventional and specific PCR for poliovirus and real time PCR for non polio EV were performed in BS and in OPV samples. Results: No genetic material of poliovirus was detected in any infant, while in 9 of them (18%) non polio EV was identified. Real time PCR for EV did not amplify poliovirus from OPV samples. Discussion: Results suggest that no post VPO viraemia detectable by molecular methods exists. Considering that real time PCR for EV does not allow to identify polio virus, no false positives of the test are expected as a result of a recent VPO vaccination. We documented presence of non polio EV in blood of healthy asymptomatic infants.

Introducción: No existen estudios que indiquen si la vacuna polio oral (VPO) produce viremia detectable mediante métodos moleculares. Una eventual viremia podría afectar el rendimiento de la RPC tiempo real para detectar enterovirus (EV) no polio, examen de creciente uso clínico en lactantes pequeños con fiebre sin foco. Objetivo: Determinar viremia post VPO en lactantes sanos, por métodos moleculares. Métodos: 50 menores de 3 meses, al momento de recibir su primera VPO se distribuyeron en forma aleatoria en 5 grupos: control, muestra de sangre pre-vacunación; grupo 1, muestra al 2° día; grupo 2, al 4° día; grupo 3, al 6° día y grupo 4, al 8° día post-vacunación. Se realizó RPC convencional específica para virus polio y RPC tiempo real para EV no polio en las muestras de sangre y en muestras de VPO. Resultados: No se identificó presencia de material genético de virus polio en lactante alguno, mientras que en 9 (18%) se identificó presencia de EV no polio. La RPC tiempo real para EV no polio no amplificó material genético a partir de las muestras de VPO. Discusión: Los resultados sugieren que no existe viremia post-VPO detectable por métodos moleculares. Considerando que la RPC tiempo real de EV no polio de uso clínico no permite identificar la presencia de virus polio, estos hallazgos indican que no existirán falsos positivos de este examen como resultado de una vacunación VPO reciente. Adicionalmente se documentó presencia de EV no polio en sangre de lactantes asintomáticos.

First description of Adenovirus, Enterovirus, Rotavirus and Torque teno virus in water samples collected from the Arroio Dilúvio, Porto Alegre, Brazil / Primeira descrição de Adenovírus, Enterovírus, Rotavírus e Torque teno vírus em amostras de água coletadas do Arroio Dilúvio, Porto Alegre, Brasil

Adenovirus (AdV), enterovirus (EV), genogroup A rotaviruses (GARV) and Torque teno virus (TTV) are non-enveloped viral agents excreted in feces and so may contaminate water bodies. In the present study, the molecular detection of these viruses was performed in samples of surface water collected from the Arroio Dilúvio, a waterstream that crosses the city of Porto Alegre, RS, Brazil, receiving great volumes of non-treated sewage from a large urban area. Sampling was performed during 2009, in three different occasions (January, April and September). The highest detection rate was observed for EV (64.28%), followed by TTV (28.57%) and AdV (21.43%). Rotaviruses were not detected. More than on kind of tested virus was detected in five (35. 71%) of 14 samples. January was the month with the highest viral detection rate, being all samples, collected in this month, positive for at least one group of tested virus. The correlation between the detection of these different viral agents and environmental factors is discussed. To the knowledge of the authors, this is the first description of viral genomes in water samples taken from the Arroio Dilúvio, Porto Alegre (Brazil).

Adenovírus (AdV), enterovírus (EV), rotavírus (GARV) e Torque teno vírus (TTV) são vírus não envelopados, excretados nas fezes, podendo, assim, contaminar corpos hídricos. No presente estudo, a detecção molecular desses agentes foi realizada em amostras de águas superficiais provenientes do Arroio Dilúvio, o qual cruza a cidade de Porto Alegre-RS, Brasil. As amostras foram coletadas em três meses diferentes (janeiro, abril e setembro) do ano de 2009. A maior taxa de detecção viral foi observada para EV (64,28%), seguida por TTV (28,57%) e AdV (21,43%). Rotavírus não foi detectado. Foi verificada presença simultânea de dois grupos virais em cinco (35,71%) das 14 amostras analisadas. Janeiro foi o mês com a maior taxa de detecção viral, sendo todas as amostras, coletadas nesse mês, positivas para, no mínimo, um grupo viral em estudo. A correlação entre a detecção desses diferentes agentes virais e os fatores ambientais é discutida. Conforme conhecimento dos autores, essa é a primeira descrição de genomas virais em amostras de água provenientes do Arroio Dilúvio, Porto Alegre, Brasil.

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil / Enterovirus e família herpesviridae como agentes etiológicos de meningites linfomonocitárias, no sul do Brasil

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

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

Performance Evaluation of Real-Q Enterovirus Quantification Kit for Enterovirus by Real-time PCR / 대한진단검사의학회지

BACKGROUND: Molecular methods have enabled rapid diagnosis of aseptic meningitis and have reduced both unnecessary therapeutic interventions and medical costs. In this study, we evaluated the analytical performance of the recently developed Real-Q Enterovirus Quantification kit (BioSewoom Inc., Korea). METHODS: We evaluated the detection limit, precision, linearity, and cross-reactivity of the Real-Q Enterovirus Quantification kit and compared it with the conventional PCR method. From March to September 2009, we tested 91 CSF specimens from patients who visited the pediatrics department of the university hospital with symptoms of aseptic meningitis or infantile sepsis, and we also tested 48 CSF specimens from patients with febrile convulsion for differential diagnosis. RESULTS: The Real-Q Enterovirus Quantification kit showed good linearity (r=0.997) within a range from 3x10(2) to 3x10(10) copies/mL, and the detection limit of the kit was 83 copies/mL. The within-run, between-run, and between-day CVs were 5.3-7.6%, 9.5-12.3%, and 11.4-13.4%, respectively. There was no cross reactivity between enteroviruses and various microorganisms. Positive results were obtained for 39.1% (25/64) of the patients suspected of aseptic meningitis and 44.4% (12/27) of the patients suspected of infantile sepsis. However, among the 48 children with febrile conversion, only 4 were positive for enterovirus. Further, the concordance with conventional PCR was high (73/74). CONCLUSIONS: The Real-Q Enterovirus Quantification kit showed excellent linearity and high reliability with a broad reportable range. It showed good detection rate when used with clinical specimens and also showed a high concordance with the conventional method. Therefore, this assay would be clinically useful not only in diagnosis of aseptic meningitis but also in differential diagnosis of infantile sepsis.

Risk Factors for Serious Bacterial Infection in Febrile Young Infants in a Community Referral Hospital

Differentiation of serious bacterial infection (SBI) from self-limiting viral illness in febrile infants younger than three months is a significant challenge for clinicians. We aimed to assess the risk factors for SBI in febrile infants. Data were obtained from 221 infants younger than three months who visited a single community referral hospital for fever and underwent a complete sepsis workup between August 2003 and July 2006. The causes of fever were febrile illness without a documented cause (FISDC, 65%), urinary tract infection (UTI, 12%), aseptic meningitis (12%), bacteremia (4%), bacterial meningitis (2%). Cerebrospinal fluid enterovirus polymerase chain reaction was positive in 28% of FISDC and 48% of aseptic meningitis cases. When UTI was excluded, the risk factors for SBI were 1) C-reactive protein (CRP) level of > or =1.87 mg/dL and 2) fevers of > or =38.9degrees C. The specificity and negative predictive values of risk factors 1) and 2) for the diagnosis of SBI were 94% and 95%, respectively. We concluded that enteroviral infection may be a major cause of febrile episodes in infants younger than three months. If UTI could be excluded, the presence of CRP levels > or =1.87 mg/dL and fevers of > or =38.9degrees C can be used as criteria to rule out SBI in these infants.

Application of a Diagnostic Method Using Reverse Transcription-PCR ELISA for the Diagnosis of Enteroviral Infections / 대한진단검사의학회지

BACKGROUND: Enteroviruses are known as major pathogen for aseptic meningitis. Although rapid diagnosis for enteroviruses is very essential to exclude bacterial infections in patients with meningitis, classical diagnostic method based on virus isolation is not practicable for timely treatment of patients due to its laborious and time-consuming procedure. Recently molecular methodologies as alternatives are routinely used for rapid and sensitive diagnosis for enteroviruses infections. METHODS: Reverse transcription (RT)-PCR ELISA kit for targeting 5'non-coding region (NCR) with highly conserved genetic identity among all genotypes of enteroviruses was introduced in this investigation. RT-PCR ELISA was evaluated about sensitivity and specificity through virus isolation using clinical specimens from patients suspected of enteroviral infections and enteroviral isolates comparing with conventional RT-PCR identifying them. RESULTS: The detection limit of the RT-PCR ELISA was up to 10-100 folds higher than virus isolation using cell culture and conventional RT-PCR. On comparison between above two methods, the detection rate of RT-PCR ELISA for clinical specimens from patients with aseptic meningitis was 7% higher than that of conventional RT-PCR targeting 5'NCR (P=0.016). CONCLUSIONS: Our results suggest that RT-PCR ELISA developed in this study could be an alternative diagnostic method for the detection of enteroviral genome with high sensitivity and specificity.

Epidemiología de los enterovirus asociados a enfermedades neurológicas / Epidemiology of enterovirus associated with neurologic diseases

El presente estudio describe los resultados de la investigación de los enterovirus humanos (HEV) mediante cultivo celular y reacción en cadena de la polimerasa y su tipificación molecular en 2167 casos de parálisis fláccida aguda, meningitis aséptica y encefalitis aguda, obtenidos entre 1991 y 1998 en la Argentina. La frecuencia de detección de HEV en parálisis fláccida aguda fue 19.5% (130/666) y de poliovirus Sabin 5.4% (36/666). La tasa de detección de HEV en los casos de meningitis fue 28.8% (231/801) y en encefalitis 3.0% (21/700). El grupo etario más afectado por las meningitis fue entre 1 y 9 años (75.3%) y en los casos de parálisis fláccida aguda, de 1 a 4 años (58%). En muestras de brotes de meningitis se identificó echovirus (E) 4, E9, E30 y E17, y en casos esporádicos virus coxsackie A (CAV) 2, B (CBV) 2 y CBV5, E7, E11, E19, E24 y E29, y enterovirus (EV) 71. Finalmente, en casos de encefalitis se detectó E4, E7 y E24. En casos de parálisis fláccida aguda se identificaron 28 serotipos distintos de enterovirus no polio. En la Argentina y en otros países latinoamericanos existe escasa información acerca de la circulación de los HEV y su relación con diversas enfermedades neurológicas. Este estudio proporciona información que puede servir como base para posteriores investigaciones.

This report describes the results of human enterovirus (HEV) detection and characterization using cell culture, polymerase chain reaction and molecular typing in 2167 samples obtained from acute flaccid paralysis, aseptic meningitis and acute encephalitis patients, from 1991 to 1998 in Argentina. HEV were isolated in 130 out of 666 cases (19.5%) and 36 out of 666 (5.4%). HEV RNA was detected in 28.8% (231/801) and 3.0% (21/700) of the patients with meningitis and encephalitis, respectively. Children with ages ranging from 1 to 9 years accounted for 75.3% of the meningitis cases and from 1 to 4 years for 58% of acute flaccid paralysis patients. Echovirus 4 (E4), E9, E30 and E17 were identified from meningitis outbreaks. Coxsackievirus A2 (CAV2), CBV2, CBV5, E7, E11, E19, E24, E29 and enterovirus 71 were recovered only from sporadic cases. Three different serotypes were identified in encephalitis patients: E4, E7 and E24. A total of 28 different serotypes of non-polio enteroviruses were detected from acute flaccid paralysis cases. The information here presented contributes to improving our knowledge about enteroviruses epidemiology in Argentina and their relationship with different neurological diseases. This study provides valuable data that could be useful to further research.

Molecular and seroepidemiologic studies of Enterovirus 71 infection in the State of Pará, Brazil

Em muitos países, o Enterovírus 71 (EV-71) família Picornaviridae é associado a doença de pé-mão e boca e doenças neurológicas agudas enquanto que no Brasil esse vírus está mais associado às últimas. O objetivo desta pesquisa foi utilizar em estudos moleculares e soroepidemiológicos, o primeiro isolamento de EV-71 obtido na região norte do Brasil. No período de janeiro de 1998 a dezembro de 2000 foram coletadas 88 amostras (44 casos de PFA) de fezes das quais, duas (2,2%) foram positivas para EV-71 (73442/PA/99). A seqüência de nucleotídeos do gen que codifica a proteína VP1 mostrou que o isolado 73442/PA/99 foi similar às cepas de EV-71 pertencentes ao grupo B- mais próxima das norte americanas. Teste de neutralização com 389 amostras de soro colhidas no período de janeiro de 1998 a novembro de 2001, de indivíduos com idade de 0 a 15 anos residentes na cidade de Belém, Estado do Pará mostrou os seguintes resultados em relação ao isolado 73442/PA/99 e ao protótipo BrCr: 207 indivíduos (53,2%) tinham anticorpos neutralizantes para ambos os vírus, 167 (42,9%) não tinham anticorpos e 15 tinham anticorpos para um dos dois vírus. Somente 20,2% das crianças com idade de 0 a 3 anos tinham anticorpos neutralizantes para EV-71, indicando que essas crianças estavam mais suscetíveis à infecção. Tanto o estudo de soroprevalência quanto o de sequenciamento da VP1 foram importantes para demonstrar a propagação e o padrão molecular do EV-71 circulante na região norte do Brasil.

Aplicación de la secuenciación nucleotídica de VP1 a la identificación de Enterovirus humanos

Se describió la introducción de un método molecular para la identificación de los Enterovirus basado en la amplificación, secuenciación y análisis filogenético de la proteína VP1. Se demostró que este método reduce grandemente el tiempo requerido para la identificación de los Enterovirus aislados y resulta de gran utilidad en la caracterización de aislamientos difíciles de tipificar, con el empleo de los reactivos inmunológicos de rutina. Por la rapidez de ejecución de la técnica reviste vital importancia su uso durante epidemias, dada la rápida determinación del agente causal.

The introduction of a mollecular method to identify the Entoviruses based on the amplification, sequencing and phylogenetic analysis of protein VP1 was described. It was proved that this method reduces significantly the time required for the identification of the isolated Entoviruses and that it is very useful in the characterization of isolates which are difficult to typify by the routine immunoloigical reagents. As it is a very fast technqiue, its use is very important during epidemics to determine the causal agent rapidly.

Análisis de un brote de meningitis viral en la provincia de Tucumán, Argentina / Analysis of an outbreak of viral meningitis in the province of Tucuman, Argentina

OBJETIVO: Confirmar la existencia de un brote de meningitis viral en 1996 en la provincia de Tucumán, Argentina, y estudiar sus características epidemiológicas. MÉTODOS: Se analizó información obtenida del Sistema Nacional de Vigilancia Epidemiológica (SINAVE) del Ministerio de Salud de Argentina para el período de 1994-1998, la cual fue provista por la Dirección de Epidemiología de dicho ministerio. Para el cálculo de incidencias se usaron estimaciones poblacionales para los años 1994-1998 realizadas por el Instituto Nacional de Estadística y Censos (INDEC) sobre la base del censo de 1991. El estudio de frecuencias se realizó mediante el análisis de tablas de contingencia de doble entrada, según el método de ji cuadrado con la corrección de Yates. Se consideró significativo el resultado cuando P < 0,05. RESULTADOS: Se confirmó la presencia de un brote de 189 casos entre el 11 de febrero y el 18 de mayo de 1996. La incidencia de casos en la provincia mostró un aumento entre 1995 y 1996 (de 0,5 a 19,3 casos por 100000 años-persona) y dicha incidencia fue significativamenrte mayor que la observada en el resto del país (19,3 frente a 2,8 casos por 100000 años-persona). El 75,1 por ciento de los casos ocurrió en niños menores de 9 años (142/189). Se detectó la presencia de Enterovirus (EV) en 65 de las 111 muestras estudiadas (58,6 por ciento). Mediante la reacción en cadena de la polimerasa (RCP) anidada con transcripción inversa se logró detectar EV en 66,3 por ciento (53/80) de los casos estudiados por este método, en comparación con solo 29,6 por ciento (24/81) de los estudiados mediante aislamiento viral. Se identificó echovirus tipo 4 en 15 (68 por ciento) en las 22 muestras tipificadas (5 por aislamiento, 3 por secuenciación y 7 por ambos métodos). Este brote demuestra la capacidad de los EV para diseminarse y producir enfermedad en la población. Durante el brote, por lo menos 56 por ciento de los casos fueron hospitalizados. CONCLUSIONES: El uso de métodos moleculares permitió el diagnóstico rápido del virus etiológico y posibilitó un mejor control del brote. El reconocimiento temprano de este podría haber evitado la mayoría de las hospitalizaciones y el uso indiscriminado de antibióticos

Objective. To confirm the occurrence of an outbreak of viral meningitis in 1996 in the province of Tucuman, Argentina, and to study the outbreak's epidemiological characteristics. Methods. We analyzed information from the National Epidemiological Surveillance System of the Ministry of Health (MOH) of Argentina for 1994­1998 that had been provided by the MOH's Bureau of Epidemiology. We calculated incidence rates using population estimates for the years 1994­1998 developed by the National Statistics and Census Institute, based on the 1991 census. We studied frequencies with contingency tables, using the chi-square method with Yates' correction. Results were considered significant when P < 0.05. Results. We confirmed the occurrence of an outbreak of 189 cases of viral meningitis between 11 February and 18 May 1996. The incidence of cases in Tucuman province increased between 1995 and 1996, from 0.5 to 19.3 cases per 100 000 person-years. That 1996 rate in Tucuman was significantly higher than what was seen in the rest of the country (2.8 cases per 100 000 person-years). Of the 189 cases, 142 of them (75.1%) occurred in children less than 9 years old. Out of 111 samples studied, 65 of them (58.6%) were enterovirus-positive. Through reverse transcription-nested polymerase chain reaction, enteroviruses were found in 66.3% (53/80) of the cases studied by this method, versus in only 29.6% (24/81) of the cases studied through viral isolation. In the 22 samples that were serotyped, echovirus type 4 was identified in 15 of them (68%): 5 by isolation, 3 by sequencing, and 7 by both methods. During the Tucuman outbreak, at least 56% of the cases were hospitalized. This viral meningitis outbreak shows the capacity of enteroviruses to spread and cause disease. Conclusions. The use of molecular methods makes it possible to rapidly diagnose the etiological virus and to better control an outbreak. Recognizing this outbreak in Tucuman sooner could have averted the majority of the hospitalizations and the indiscriminate use of antibiotics.

Detection of enteroviral sequence in endomyocardial tissues from patients with cardiac diseases in northern Brazil

No presente trabalho, são mostrados resultados de um estudo piloto direcionado à detecção de seqüências de enterovirus em tecido cardíaco obtido a partir de biópsias endomiocárdicas de pacientes com doenças cardíacas na região Amazônica. Seis amostras coletadas de três pacientes foram analisadas por RT-PCR obtendo-se três espécimes positivos e três negativos. Esses achados preliminares sugerem a participação dos enterovirus na etiologia de doenças cardíacas, principalmente miocardites, e justificam estudos mais amplos nesse assunto.

In the present report we describe the results from a pilot study aimed at detecting enterovirus sequence in cardiac tissues, obtained through endomyocardial biopsies, from patients suffering from cardiac diseases in the Amazon region. Six samples that were collected from three patients were analysed by RT-PCR showing 3 positive and 3 negative results. These preliminary findings suggest the participation of enteroviruses in the etiology of cardiac diseases, mainly myocarditis, and warrant further and broader local studies on this subject.

Caracterización molecular de enterovirus no-polio procedentes de casos con parálisis residual / Molecular characterization of non-polio enteroviruses from residual paralysis cases. Colombia, 1992 - 1995

Las parálisis fláccidas agudas (PFA) tienen una amplia variedad de orígenes y de agentes causales: físicos (traumas), fisiopatológicos (Accidente cerebro vascular (ACV), tóxicos (drogas o químicos) e infecciosos (bacterias y virus). Entre estos últimos, el virus salvaje de la poliomielitis y el enterovirus 71 (EV71), son los agentes virales más frecuentes. Con la no-detección de poliovirus salvaje autóctono como agente causal de enfermedad paralítica en Colombia desde junio de 1991 y aislamientos de virus No-polio en el 20.84 por ciento del total de casos de PFA notificados anualmente, se quiso conocer el papel que jugan los enterovirus en la incidencia de parálisis fláccida Aguda y la dinámica de circulación y distribución de los mismos en Colombia, para lo cual, se revisó la base de datos epidemiológicos y clínicos de los casos notificados al programa de erradicación de la poliomielitis en Colombia a partir del 1º de enero de 1992 al 31 de diciembre de 1995. Se clasificaron los casos con base en la presencia de parálisis residual y la entidad clínica de descarte según valoración y clasificación realizada por el Grupo de control de patologías del Ministerio de Salud, el Programa Ampliado de Inmunizaciones (PAI), la Organización Panamericana de la Salud y la Organización Mundial de la Salud (OPS/OMS). Durante estos cuatro años, el Sistema de Vigilancia Epidemiológica de las parálisis fláccideas, notificó 856 casos en menores de 15 años, de los cuales, 346 (40.42 por ciento) presentaron paralisis residual, 331 (95.6 por ciento) tuvieron muestras de heces para estudio virológico. Se seleccionaron los casos con estudio virológico para Enterovirus (incluyendo poliovirus) y se encontró que las patologías más frecuentes fueron (Síndrome de Guillaín Barré, neuropatía periférica, encefalitis y meningitis virales, hemiplejía infantil aguda, esclerosis múltiple, mielitis transversa y mielopatías, miositis, polimiositis, monoparesia y dermatomiositis. De estos, 69 casos (20.84 por ciento) tuvieron aislamiento de virus No polio. En 16 casos (4.8 por ciento el aislamiento fue un poliovirus vacunal, 5 de los cuales (1.2 por ciento) se asociaron a poliomielitis paralítica post-vacunal. Se realizó identificación de serotipos mediante neutralización con mezclas de antisueros anti-enterovirus de Lim & Benyesh-Melnick -LBM- (40,41) y caracterización molecular mediante reacción en cadena de la polimerasa -PCR- utilizando primers complementarios a la región VP1 del...

Neuropatía epidémica. Hipótesis etiopatogénica / Epidemic neuropathy. Etiopathogenic hypothesis

Durante la epidemia de neuropatía ocurrida en Cuba en los años 1992/1994 se aislaron del líquido cefalorraquídeo de pacientes agentes virales relacionados antigénicamente con los virus Coxsackie. Para establecer una función de estos virus en la etiopatogenia de la enfermedad se seleccionaron las cepas 47/93 IPK identificada como Coxsackie A9 y la cepa 44/93 IPK de efecto citopático ligero (ECP-L) y se realizó un estudio de sus características antigénicas mediante Western Blot. Se comprobó la relación antigénica entre ambas cepas y se demostró la ausencia de proteínas estructurales en su forma nativa en los agentes de ECP-L. A partir de estos resultados se plantea la posibilidad de que la persistencia sea un mecanismo por el cual estos virus participen en la etiopatogenia de la neuropatía epidémica en Cuba

Caracterización genómica de la cepa cubana de efecto citopático ligero / Genomic characterization of the Cuban strain of light cytopathic effect

Durante la epidemia de neuropatía ocurrida en Cuba en los años 1992/1994 se aislaron del líquido cefalorraquídeo de pacientes agentes virales relacionados antigénicamente con los virus Coxsackie. Para establecer una función de estos virus en la etiopatogenia de la enfermedad se seleccionaron las cepas 47/93 IPK identificada como Coxsackie A9 y la cepa 44/93 IPK de efecto citopático ligero (ECP-L) y se realizó un estudio de sus características antigénicas mediante Western Blot. Se comprobó la relación antigénica entre ambas cepas y se demostró la ausencia de proteínas estructurales en su forma nativa en los agentes de ECP-L. A partir de estos resultados se plantea la posibilidad de que la persistencia sea un mecanismo por el cual estos virus participen en la etiopatogenia de la neuropatía epidémica en Cuba