RESUMO
BACKGROUND: Infectious diarrhea is still a major problem in public health, especially in children under 5 years of age. The identification of the etiologic agent is important for the clinical management of the diarrhea episode and, from the epidemiological point of view, to implement control measures. AIM: To determine the presence of gastrointestinal pathogens in children under five years of age with diarrhea in a Chilean rotavirus surveillance center. METHODS: Observational study in children under five years of age who were hospitalized for diarrhea at the Dr. Luis Calvo Mackenna Hospital from December 2015 to December 2019. Molecular detection was performed using the FilmArray gastrointestinal (FilmArray GI®) panel. RESULTS: We analyzed 493 diarrheal stool samples of children, 427 samples (87%) were positive and 66 samples (13%) were negative. Of positive samples, 174 samples (41%) and 253 samples (59%) were positive for one or more pathogen, respectively. In children under one year and the group between one and four years there was a predominance of infections caused by enteric virus. Rotavirus and norovirus were the most common virus in both age groups. The most frequent bacteria were EPEC (27%), C. difficile (17%), EAEC (14%) and Campylobacter (9%). In parasites, Giardia lamblia and Cryptosporidium were identified, in 3% and 1% of the total samples, respectively. CONCLUSIONS: The molecular detection system used allowed an increase in the detection of enteropathogens in children under five years of age. The information generated by this type of surveillance could help to characterize the episodes of diarrhea in the population and might be a tool to technically advise the authorities in the decision-making process for the implementation of control measures.
Assuntos
Clostridioides difficile , Criptosporidiose , Cryptosporidium , Infecções por Rotavirus , Rotavirus , Animais , Criança , Pré-Escolar , Chile/epidemiologia , Diarreia/epidemiologia , Fezes , Hospitais , Humanos , Lactente , Rotavirus/genética , Infecções por Rotavirus/epidemiologia , Vigilância de Evento SentinelaRESUMO
INTRODUCCIÓN: Las diarreas de causa infecciosa son un problema de salud pública, especialmente en niños bajo los cinco años. La identificación de los agentes etiológicos puede ser relevante para el manejo del cuadro clínico y, desde el punto de vista epidemiológico, para la implementación de medidas de control. OBJETIVO: Determinar la presencia de patógenos entéricos en niños bajo los cinco años que se hospitalizaron por diarrea aguda en uno de los centros centinelas de la red de vigilancia de rotavirus en Chile. PACIENTES Y MÉTODOS: Estudio observacional en niños menores de cinco años que se internaron por cuadros de diarrea en el Hospital Dr. Luis Calvo Mackenna, durante diciembre del 2015 a diciembre del 2019, el que forma parte de la red de vigilancia de rotavirus del Ministerio de Salud de Chile. Las muestras fecales se analizaron mediante un test molecular, FilmArray GI® panel, que permite la detección de 22 patógenos entéricos virales, bacterianos y parasitarios. RESULTADOS: Se analizaron 493 muestras fecales de niños con episodios de diarrea infecciosa, detectando al menos un patógeno en 427 muestras (87%). De estas muestras positivas, se detectó solo un patógeno en 174 muestras (41%) y dos o más patógenos en 253 muestras (59%). En el grupo de niños bajo un año y el grupo entre uno y cuatro años hubo un predominio de infecciones causadas por virus gastroentéricos, siendo rotavirus y norovirus los virus más detectados en ambos grupos de edad. Las bacterias más frecuentes fueron EPEC (27%), C. difficile (17%), EAEC (14%) y Campylobacter (9%). Respecto a los parásitos, se identificó Giardia lamblia y Cryptosporidium, en el 3 y 1% del total de las muestras, respectivamente. CONCLUSIÓN: La detección molecular utilizada permitió detectar un alto número de enteropatógenos en niños bajo los cinco años. La información generada por este tipo de vigilancia, podría ayudar a caracterizar en la población los episodios de diarrea causados por los principales patógenos entéricos y podría ser una herramienta para asesorar técnicamente a las autoridades en la toma de decisión para la implementación de medidas de control contra estos patógenos.
BACKGROUND: Infectious diarrhea is still a major problem in public health, especially in children under 5 years of age. The identification of the etiologic agent is important for the clinical management of the diarrhea episode and, from the epidemiological point of view, to implement control measures. AIM: To determine the presence of gastrointestinal pathogens in children under five years of age with diarrhea in a Chilean rotavirus surveillance center. METHODS: Observational study in children under five years of age who were hospitalized for diarrhea at the Dr. Luis Calvo Mackenna Hospital from December 2015 to December 2019. Molecular detection was performed using the FilmArray gastrointestinal (FilmArray GI®) panel. RESULTS: We analyzed 493 diarrheal stool samples of children, 427 samples (87%) were positive and 66 samples (13%) were negative. Of positive samples, 174 samples (41%) and 253 samples (59%) were positive for one or more pathogen, respectively. In children under one year and the group between one and four years there was a predominance of infections caused by enteric virus. Rotavirus and norovirus were the most common virus in both age groups. The most frequent bacteria were EPEC (27%), C. difficile (17%), EAEC (14%) and Campylobacter (9%). In parasites, Giardia lamblia and Cryptosporidium were identified, in 3% and 1% of the total samples, respectively. CONCLUSIONS: The molecular detection system used allowed an increase in the detection of enteropathogens in children under five years of age. The information generated by this type of surveillance could help to characterize the episodes of diarrhea in the population and might be a tool to technically advise the authorities in the decision-making process for the implementation of control measures.
Assuntos
Humanos , Animais , Lactente , Pré-Escolar , Criança , Infecções por Rotavirus , Clostridioides difficile , Rotavirus , Criptosporidiose , Cryptosporidium , Infecções por Rotavirus/epidemiologia , Chile/epidemiologia , Rotavirus/genética , Vigilância de Evento Sentinela , Diarreia/epidemiologia , Fezes , HospitaisRESUMO
Andes virus (ANDV) is the etiological agent of hantavirus cardiopulmonary syndrome in Chile. In this study, we evaluated the profile of the pro-inflammatory cytokines IL-1ß, IL-12p70, IL-21, TNF-α, IFN-γ, IL-10 and IL-6 in serum samples of ANDV-infected patients at the time of hospitalization. The mean levels of circulating cytokines were determined by a Bead-Based Multiplex assay coupled with Luminex detection technology, in order to compare 43 serum samples of healthy controls and 43 samples of ANDV-infected patients that had been categorized according to the severity of disease. When compared to the controls, no significant differences in IL-1ß concentration were observed in ANDV-infected patients (p = 0.9672), whereas levels of IL-12p70 and IL-21 were significantly lower in infected cases (p = <0.0001). Significantly elevated levels of TNF-α, IFN-γ, IL-10, and IL-6 were detected in ANDV-infected individuals (p = <0.0001, 0.0036, <0.0001, <0.0001, respectively). Notably, IL-6 levels were significantly higher (40-fold) in the 22 patients with severe symptoms compared to the 21 individuals with mild symptoms (p = <0.0001). Using multivariate regression models, we show that IL-6 levels has a crude OR of 14.4 (CI: 3.3-63.1). In conclusion, the serum level of IL-6 is a significant predictor of the severity of the clinical outcome of ANDV-induced disease.
Assuntos
Progressão da Doença , Síndrome Pulmonar por Hantavirus/sangue , Síndrome Pulmonar por Hantavirus/epidemiologia , Interleucina-6/sangue , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Criança , Pré-Escolar , Chile/epidemiologia , Citocinas/sangue , Feminino , Orthohantavírus , Síndrome Pulmonar por Hantavirus/fisiopatologia , Humanos , Lactente , Recém-Nascido , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Fatores de Risco , Índice de Gravidade de Doença , Distribuição por Sexo , Adulto JovemRESUMO
BACKGROUND: Andes virus (ANDV) is the sole etiologic agent of hantavirus cardiopulmonary syndrome (HCPS) in Chile, with a fatality rate of about 35%. Individual host factors affecting ANDV infection outcome are poorly understood. In this case-control genetic association analysis, we explored the link between single-nucleotide polymorphisms (SNPs) rs12979860, rs8099917 and rs1800629 and the clinical outcome of ANDV-induced disease. The SNPs rs12979860 and rs8099917 are known to play a role in the differential expression of the interleukin 28B gene (IL28B), whereas SNP rs1800629 is implicated in the expression of tumor necrosis factor α gene (TNF-α). METHODS: A total of 238 samples from confirmed ANDV-infected patients collected between 2006 and 2014, and categorized according to the severity of the disease, were genotyped for SNPs rs12979860, rs8099917, and rs1800629. RESULTS: Analysis of IL28B SNPs rs12979860 and rs8099917 revealed a link between homozygosity of the minor alleles (TT and GG, respectively), displaying a mild disease progression, whereas heterozygosity or homozygosity for the major alleles (CT/CC and TG/TT, respectively) in both IL28B SNPs is associated with severe disease. No association with the clinical outcome of HCPS was observed for TNF-α SNP rs1800629 (TNF -308G>A). CONCLUSIONS: The IL28B SNPs rs12979860 and rs8099917, but not TNF-α SNP rs1800629, are associated with the clinical outcome of ANDV-induced disease, suggesting a possible link between IL28B expression and ANDV pathogenesis.
Assuntos
Infecções por Hantavirus/genética , Infecções por Hantavirus/patologia , Interleucinas/genética , Orthohantavírus/isolamento & purificação , Polimorfismo de Nucleotídeo Único , Índice de Gravidade de Doença , Fator de Necrose Tumoral alfa/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Criança , Pré-Escolar , Chile , Feminino , Estudos de Associação Genética , Técnicas de Genotipagem , Infecções por Hantavirus/imunologia , Humanos , Lactente , Recém-Nascido , Interferons , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
The focus assay is currently the most commonly used technique for hantavirus titer determination. This method requires an incubation time of between 5 and 11 days to allow the appearance of foci after several rounds of viral infection. The following work presents a rapid Andes virus (ANDV) titration assay, based on viral nucleocapsid protein (N) detection in infected cells by flow cytometry. To this end, an anti-N monoclonal antibody was used that was developed and characterized previously. ANDV N could be detected as early as 6 h post-infection, while viral release was not observed until 24-48 h post-infection. Given that ANDV detection was performed during its first round of infection, a time reduction for titer determination was possible and provided results in only two days. The viral titer was calculated from the percentage of N positive cells and agreed with focus assay titers. Furthermore, the assay was applied to quantify the inhibition of ANDV cell entry by patient sera and by preventing endosome acidification. This novel hantavirus titration assay is a highly quantitative and sensitive tool that facilitates infectivity titration of virus stocks, rapid screening for antiviral drugs, and may be further used to detect and quantify infectious virus in human samples.
Assuntos
Citometria de Fluxo/métodos , Viabilidade Microbiana , Orthohantavírus/isolamento & purificação , Orthohantavírus/fisiologia , Carga Viral/métodos , Animais , Anticorpos Monoclonais , Anticorpos Antivirais , Chlorocebus aethiops , Proteínas do Nucleocapsídeo/análise , Sensibilidade e Especificidade , Fatores de Tempo , Células VeroRESUMO
OBJETIVO: las diarreas son una de las principales causas de morbilidad y mortalidad en el mundo. En Chile, su vigilancia es una prioridad, muy en particular las causadas por rotavirus para conocer su epidemiología, incluidos los serotipos circulantes y detectar brotes oportunamente. MÉTODOS: se implementó un sistema de vigilancia en 8 hospitales pedißtricos, considerados sitio centinela por cubrir 62 por ciento de la población menor de 5 años de las regiones Metropolitana, Valparaíso y Bio-Bio. Se utilizaron las definiciones de caso estandarizadas por la Organización Mundial de la Salud. Las muestras fueron analizadas en los laboratorios del Instituto de Salud Pública con la técnica inmunoensayo enzimßtico en el formato de ELISA. El sistema se basa en un formulario electrónico en línea, sobre plataforma de programación PHP versión 5.2.6, diseñado para creación de páginas web dinßmicas, accesible mediante clave o password de los terminales del centro centinela, de la Secretaria Regional Ministerial de Salud e Instituto de Salud Pública, donde se introducen las variables semanal y mensualmente de forma definitiva. El anßlisis se realiza en el programa estadístico Epinfo-2000. RESULTADOS: en 8 meses de vigilancia, se estudiaron 599 muestras de deposiciones, 164 positivas a rotavirus (27,4 por ciento), tasa de 0,2 por 1 000 habitantes. Al sexo masculino pertenece 56 por ciento, el menor de 2 años es el más afectado (69 por ciento). CONCLUSIONES: la implementación del sistema de vigilancia centinela para rotavirus constituye una herramienta útil para conocer el comportamiento de esta enfermedad y contribuir a la toma de decisiones por los programas nacionales de salud(AU)
OBJECTIVE: diarrheas are one of the main causes of morbidity and mortality worldwide. In Chile, diarrhea surveillance is a priority, particularly those caused by rotavitus, in order to find out their epidemiology including circulating serotypes and to promptly detect possible outbreaks. METHODS: a surveillance system was implemented in 8 pediatric hospitals considered as sentinel sites because they cover 62 percent of the under 5 years-old population from Metropolitana, Valparaiso and Bio-Bio regions. The standardized case definitions of the World Health Organization were used. The sample analysis was based on ELISA method in the Public Health Institute laboratories. The surveillance system is based on an online electronic form supported on PHP platform, 5.2.6 version, designed to create dynamic and accessible webpages through passwords from the sentinel center terminals of the Regional Ministerial Department of Health and the Public Health Institute; the variables are weekly and monthly introduced. The statistical analysis is carried out through Epinfo-2000. RESULTS: five hundred and ninety nine fecal samples were analyzed in eight months; 164 of them were positive to rotavirus (27.4 percent) for a rate of 0.2 per 1000 pop. Fifty six percent were males whereas under 2 years-old group was the most affected (69 percent). CONCLUSIONS: the implementation of the sentinel rotavirus surveillance system is a useful tool for the analysis of this disease behavior and for decision-making in the health national programs(AU)
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Monitoramento Epidemiológico/normas , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/prevenção & controleRESUMO
OBJETIVO: las diarreas son una de las principales causas de morbilidad y mortalidad en el mundo. En Chile, su vigilancia es una prioridad, muy en particular las causadas por rotavirus para conocer su epidemiología, incluidos los serotipos circulantes y detectar brotes oportunamente. MÉTODOS: se implementó un sistema de vigilancia en 8 hospitales pedißtricos, considerados sitio centinela por cubrir 62 por ciento de la población menor de 5 años de las regiones Metropolitana, Valparaíso y Bio-Bio. Se utilizaron las definiciones de caso estandarizadas por la Organización Mundial de la Salud. Las muestras fueron analizadas en los laboratorios del Instituto de Salud Pública con la técnica inmunoensayo enzimßtico en el formato de ELISA. El sistema se basa en un formulario electrónico en línea, sobre plataforma de programación PHP versión 5.2.6, diseñado para creación de páginas web dinßmicas, accesible mediante clave o password de los terminales del centro centinela, de la Secretaria Regional Ministerial de Salud e Instituto de Salud Pública, donde se introducen las variables semanal y mensualmente de forma definitiva. El anßlisis se realiza en el programa estadístico Epinfo-2000. RESULTADOS: en 8 meses de vigilancia, se estudiaron 599 muestras de deposiciones, 164 positivas a rotavirus (27,4 por ciento), tasa de 0,2 por 1 000 habitantes. Al sexo masculino pertenece 56 por ciento, el menor de 2 años es el más afectado (69 por ciento). CONCLUSIONES: la implementación del sistema de vigilancia centinela para rotavirus constituye una herramienta útil para conocer el comportamiento de esta enfermedad y contribuir a la toma de decisiones por los programas nacionales de salud.
OBJECTIVE: diarrheas are one of the main causes of morbidity and mortality worldwide. In Chile, diarrhea surveillance is a priority, particularly those caused by rotavitus, in order to find out their epidemiology including circulating serotypes and to promptly detect possible outbreaks. METHODS: a surveillance system was implemented in 8 pediatric hospitals considered as sentinel sites because they cover 62 percent of the under 5 years-old population from Metropolitana, Valparaiso and Bio-Bio regions. The standardized case definitions of the World Health Organization were used. The sample analysis was based on ELISA method in the Public Health Institute laboratories. The surveillance system is based on an online electronic form supported on PHP platform, 5.2.6 version, designed to create dynamic and accessible webpages through passwords from the sentinel center terminals of the Regional Ministerial Department of Health and the Public Health Institute; the variables are weekly and monthly introduced. The statistical analysis is carried out through Epinfo-2000. RESULTS: five hundred and ninety nine fecal samples were analyzed in eight months; 164 of them were positive to rotavirus (27.4 percent) for a rate of 0.2 per 1000 pop. Fifty six percent were males whereas under 2 years-old group was the most affected (69 percent). CONCLUSIONS: the implementation of the sentinel rotavirus surveillance system is a useful tool for the analysis of this disease behavior and for decision-making in the health national programs.
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/prevenção & controle , Monitoramento Epidemiológico/normasRESUMO
Andes virus (ANDV) is the predominant etiologic agent of hantavirus cardiopulmonary syndrome (HCPS) in southern South America. In Chile, serologically confirmed human hantavirus infections have occurred throughout a wide latitudinal distribution extending from the regions of Valparaíso (32 to 33 degrees S) to Aysén (46 degrees S) in southern Patagonia. In this study, we found seropositive rodents further north in the Coquimbo region (30 degrees S) in Chile. Rodent seroprevalence was 1.4%, with Oligoryzomys longicaudatus displaying the highest seroprevalence (5.9%), followed by Abrothrix longipilis (1.9%) and other species exhibiting
Assuntos
Variação Genética , Infecções por Hantavirus/veterinária , Infecções por Hantavirus/virologia , Orthohantavírus/classificação , Orthohantavírus/isolamento & purificação , Filogenia , Animais , Chile/epidemiologia , Feminino , Geografia , Orthohantavírus/genética , Infecções por Hantavirus/epidemiologia , Humanos , Masculino , Epidemiologia Molecular , Dados de Sequência Molecular , RNA Viral/genética , Recombinação Genética , Roedores , Análise de Sequência de DNA , Estudos SoroepidemiológicosRESUMO
In the present work we identified B-cell epitopes recognized by sera of humans and rodents naturally infected with Andes virus, a hantavirus present in Chile and Argentina. Analysis of patient and rodent sera with overlapping peptides revealed 21 human and rodent epitopes on the three structural proteins. Whereas in the nucleoprotein the region comprising aa 248-260 was shown to be the key determinant of human sera, the major antigenic site of rodent antibody reactivity is located at aa 326-338. In G1, the main epitope recognized by human sera was mapped to aa 14-26, while rodent antibodies bound predominantly to aa 599-611. In contrast, humans and mice had strong responses to three regions in G2 (aa 691-703, aa 918-930, aa 955-967), of which the last two are associated with neutralization of Hantaan virus. This insight affords important information for the development of immunotherapies for the acute phase of hantavirus cardiopulmonary syndrome.
Assuntos
Mapeamento de Epitopos , Epitopos de Linfócito B/imunologia , Infecções por Hantavirus/imunologia , Orthohantavírus/imunologia , Doenças dos Roedores/imunologia , Proteínas Estruturais Virais/imunologia , Adolescente , Adulto , Sequência de Aminoácidos , Animais , Anticorpos Antivirais/sangue , Linfócitos B/imunologia , Epitopos de Linfócito B/química , Feminino , Infecções por Hantavirus/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Testes de Neutralização , Doenças dos Roedores/virologia , Proteínas Estruturais Virais/químicaRESUMO
We report here the complete genomic sequence of the Chilean human isolate of Andes virus CHI-7913. The S, M, and L genome segment sequences of this isolate are 1,802, 3,641 and 6,466 bases in length, with an overall GC content of 38.7%. These genome segments code for a nucleocapsid protein of 428 amino acids, a glycoprotein precursor protein of 1,138 amino acids and a RNA-dependent RNA polymerase of 2,152 amino acids. In addition, the genome also has other ORFs coding for putative proteins of 34 to 103 amino acids. The encoded proteins have greater than 98% overall similarity with the proteins of Andes virus isolates AH-1 and Chile R123. Among other sequenced Hantavirus, CHI-7913 is more closely related to Sin Nombre virus, with an overall protein similarity of 92%. The characteristics of the encoded proteins of this isolate, such as hydrophobic domains, glycosylation sites, and conserved amino acid motifs shared with other Hantavirus and other members of the Bunyaviridae family, are identified and discussed.
Assuntos
Genoma Viral , Orthohantavírus/genética , Homologia de Sequência do Ácido Nucleico , Sequência de Aminoácidos/genética , Criança , Orthohantavírus/química , Humanos , Dados de Sequência Molecular , Proteínas do Nucleocapsídeo/genética , Reação em Cadeia da Polimerase , RNA Viral/genéticaRESUMO
We report here the complete genomic sequence of the Chilean human isolate of Andes virus CHI-7913. The S, M, and L genome segment sequences of this isolate are 1,802, 3,641 and 6,466 bases in length, with an overall GC content of 38.7 percent. These genome segments code for a nucleocapsid protein of 428 amino acids, a glycoprotein precursor protein of 1,138 amino acids and a RNA-dependent RNA polymerase of 2,152 amino acids. In addition, the genome also has other ORFs coding for putative proteins of 34 to 103 amino acids. The encoded proteins have greater than 98 percent overall similarity with the proteins of Andes virus isolates AH-1 and Chile R123. Among other sequenced Hantavirus, CHI-7913 is more closely related to Sin Nombre virus, with an overall protein similarity of 92 percent. The characteristics of the encoded proteins of this isolate, such as hydrophobic domains, glycosylation sites, and conserved amino acid motifs shared with other Hantavirus and other members of the Bunyaviridae family, are identified and discussed
Assuntos
Humanos , Criança , Sequência de Aminoácidos , Genoma Viral , Homologia de Sequência do Ácido Nucleico , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , RNA ViralRESUMO
We isolated Andes virus (formal name: Andes virus [ANDV], a species in the genus Hantavirus), from serum of an asymptomatic 10-year-old Chilean boy who died 6 days later of hantavirus pulmonary syndrome (HPS). The serum was obtained 12 days after his grandmother died from HPS and 2 days before he became febrile. No hantavirus immunoglobulin (Ig) G or IgM antibodies were detected in the serum sample. After three blind passages, ANDV antigens were detected in Vero E6 cells by immunofluorescence assay and enzyme-linked immunosorbent assay, and ANDV RNA was detected by reverse transcription-polymerase chain reaction. A fragment of the virus genome showed 96.2% nucleotide identity with that of prototype ANDV. To our knowledge, this is the first isolation of any agent of hemorrhagic fever with HPS from a human and the first such isolation of hantavirus before symptoms of that syndrome or HPS began.
Assuntos
Infecções por Hantavirus/virologia , Orthohantavírus/isolamento & purificação , Animais , Criança , Chile/epidemiologia , Chlorocebus aethiops , Evolução Molecular , Evolução Fatal , Feminino , Imunofluorescência , Orthohantavírus/genética , Orthohantavírus/imunologia , Infecções por Hantavirus/epidemiologia , Infecções por Hantavirus/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Tempo , Células VeroRESUMO
The aim of this study was to determine anti HTLV-I antibody titers in seropositive symptomatic and asymptomatic infected subjects. One hundred seven infected subjects (47 with spastic paraparesis and 60 asymptomatic) were studied. HTLV-I antibodies were determined using indirect immunofluorescence in cells infected with the retrovirus. The mean titer was 1/234 in asymptomatic subjects and 1/2138 in symptomatic patients (p<0,001). These results suggest an association between HTLV-I antibody titers and clinical stage of infected subjects
