ABSTRACT
We assessed maternal and infant cytomegalovirus (CMV) infection in Colombia. Maternal serum was tested for CMV immunoglobulin G antibodies at a median of 10 (interquartile range: 8-12) weeks gestation (n = 1501). CMV DNA polymerase chain reaction was performed on infant urine to diagnose congenital (≤21 days of life) and postnatal (>21 days) infection. Maternal CMV seroprevalence was 98.1% (95% confidence interval [CI]: 97.5%-98.8%). Congenital CMV prevalence was 8.4 (95% CI: 3.9%-18.3%; 6/711) per 1000 live births. Among 472 infants without confirmed congenital CMV infection subsequently tested at age 6 months, 258 (54.7%, 95% CI: 50.2%-59.1%) had postnatal infection.
Subject(s)
Antibodies, Viral/blood , Cytomegalovirus Infections/epidemiology , Cytomegalovirus Infections/immunology , Cytomegalovirus/immunology , Pregnancy Complications, Infectious/virology , Adult , Child, Preschool , Colombia/epidemiology , Cytomegalovirus/genetics , Cytomegalovirus Infections/congenital , Cytomegalovirus Infections/urine , DNA, Viral/urine , Female , Gestational Age , Humans , Immunoglobulin G/blood , Infant , Mothers , Pregnancy , Pregnancy Complications, Infectious/epidemiology , Pregnancy Complications, Infectious/immunology , Saliva/virology , Seroepidemiologic StudiesABSTRACT
Hepatitis E virus (HEV) is one of the main causes of acute viral hepatitis of enteric transmission. HEV has been detected in environmental samples in several countries from Europe and Asia, constituting a risk factor for waterborne infection. In Colombia, HEV has been identified in samples obtained from patients as well as from swine, but no environmental studies have been carried out. To determine if HEV is present in environmental waters, samples from the main source of drinking water plant and of wastewater system of eight municipalities and two villages of Antioquia state (North West Colombia), were collected between December 2012 and April 2014. The HEV genome was detected by RT-PCR in 23.3% (7/30) of the samples from the main source of drinking water plants and in 16.7% (5/30) from sewage. Viral concentrates obtained from three positive sewage samples were used to inoculate HepG2 cell cultures that were followed for one month; however, the viral genome was not detected in any cell culture. This study demonstrates the circulation of HEV in both source of drinking water plants and wastewater in Antioquia state, Colombia. The presence of HEV in environmental waters could be a risk for waterborne transmission in this population. The findings of the present study, together with the evidence of HEV circulation in human and swine in Colombia, should be consider by public health authorities for the development of surveillance programs and the inclusion of HEV infection diagnosis in the guidelines of viral hepatitis in the country. This is the first report of HEV in environmental samples in Colombia and the second one in Latin America.
Subject(s)
Hepatitis E virus/isolation & purification , Hepatitis E/epidemiology , Hepatitis E/virology , Water Microbiology , Animals , Colombia/epidemiology , Environmental Monitoring , Genome, Viral , Geography , Hepatitis E virus/genetics , Humans , Phylogeny , RNA, Viral , Sewage/virology , Viral LoadABSTRACT
INTRODUCTION: Hepatitis A virus (HAV) is an important pathogen, typically transmitted via the faecal-oral route. The epidemiology of the infection is directly related to drinking water access and adequate disposal of sewage water. OBJECTIVE: To determine the presence and identify the genotype of HAV in environmental samples from eight municipalities and two villages in Antioquia, northwestern Colombia. MATERIALS AND METHODS: Three serial samplings were done between December, 2012, and April, 2014. Water samples were obtained from drinking water plants prior to treatment, as well as from the main reserve of wastewater in each municipality included in the study. Viral concentrations for the two types of sample sources were determined by filtration/tangential ultrafiltration and polyethyleneglycol plus flocculation with skimmed milk, respectively. Total ARN was subsequently obtained from each sample and the VP3-VP1 region amplified for detection of the viral genome. The genotype was determined by amplification of the VP1-2B region. RESULTS: The HAV genome was detected in samples from drinking water plants at Puerto Berrío, Frontino and Nutibara, and in wastewater samples from the municipalities of Arboletes, Zaragoza and Venecia. HAV subgenotype IA was identified using phylogenetic analysis. CONCLUSION: In this study, HAV was identified in 6.6% of the samples from drinking water plants and 13.3% of wastewater samples. This is the first report of HAV subgenotype IA circulating in environmental samples from Antioquia.
Subject(s)
Environmental Microbiology , Genome, Viral/genetics , Hepatitis A virus/genetics , Hepatitis A/epidemiology , Colombia/epidemiology , Genotype , Hepatitis A/virology , Humans , Phylogeny , RNA, Viral/geneticsABSTRACT
INTRODUCTION: Since drinking water can be a vehicle for the transmission of pathogens, the detection of enteric viruses in these water samples is essential to establish the appropriate measures to control and prevent associated diseases. OBJECTIVE: To analyze the results obtained for enteric viruses in water samples for human consumption received at the Colombian Instituto Nacional de Salud and establish their association with the data on water quality in Colombian municipalities. MATERIALS AND METHODS: We conducted a descriptive-retrospective analysis of the results obtained in the detection of rotavirus, enterovirus, hepatitis A virus and adenovirus in water samples received for complementary studies of enteric hepatitis, acute diarrheal disease and foodborne diseases. Data were correlated with the results of water quality surveillance determined by the national human consumption water quality index (IRCA). RESULTS: Of the 288 samples processed from 102 Colombian municipalities, 50.7% were positive for viruses: 26.73% for hepatitis A virus, 20.48% for enterovirus and rotavirus and 18.05% for adenovirus. Viruses were detected in 48.26% of non-treated water samples and in 45.83% of treated water samples. The IRCA index showed no correlation with the presence of viruses. CONCLUSIONS: The presence of viruses in water represents a public health risk and, therefore, the prevention of virus transmission through water requires appropriate policies to reinforce water supply systems and improve epidemiological surveillance.
Subject(s)
Adenoviridae/pathogenicity , Enterovirus Infections/microbiology , Enterovirus/chemistry , Viruses/immunology , Water Microbiology , Adenoviridae/chemistry , Colombia , Enterovirus/immunology , Enterovirus/metabolism , Enterovirus Infections/epidemiology , Enterovirus Infections/metabolism , Humans , Retrospective Studies , Viruses/metabolism , Water SupplyABSTRACT
Introducción. El virus de la hepatitis E se ha convertido en un problema de salud pública, especialmente en los países en desarrollo. Se conocen cuatro genotipos en mamíferos, de los cuales el G3 se ha encontrado en hepatitis autóctonas en países y regiones con gran población de cerdos, y el G1 se ha asociado a muertes maternas. Objetivo. Determinar la infección simultánea con el virus de la hepatitis E y sus genotipos circulantes en Colombia en 1.097 sueros utilizando los marcadores serológicos de los virus de las hepatitis A, B y C. Materiales y métodos. Se seleccionaron 1.097 sueros provenientes de diferentes municipios de Colombia, conservados en el Laboratorio de Virología del Instituto Nacional de Salud. Se determinaron los anticuerpos IgG e IgM anti-hepatitis E. A los positivos se les amplificó el genoma viral mediante reacción en cadena de la polimerasa convencional. Los productos se secuenciaron y analizaron filogenéticamente y se los comparó con las secuencias del ORF2 registradas en el GenBank. Resultados. Se identificaron 278 sueros positivos para IgG anti-hepatitis E, 62 para IgM y 64 para ambos marcadores. La infección simultánea con los virus de la hepatitis E y la hepatitis A determinada por IgG anti-hepatitis E fue de 33,6 % y por IgM anti-hepatitis E fue de 16,1 %; la infección simultánea por los virus de la hepatitis E y B fue de 23,4 % y 8,1 %, y por los virus de la hepatitis E y C fue de 35,4 % y 5,83 %, respectivamente. De las 52 muestras positivas en la reacción en cadena de la polimerasa convencional, nueve secuencias se agruparon como genotipo 3a de origen porcino, cepa norteamericana. Conclusiones. La mayor seropositividad se registró para las hepatitis A y E. La frecuencia de la infección simultánea con el virus de la hepatitis E y otros virus hepatótropos indica que este patógeno puede ser más frecuente de lo esperado. La circulación del genotipo 3a implica que esta enfermedad puede presentarse en forma de brote y de zoonosis en Colombia.
Introduction: Hepatitis E virus has emerged as a public health problem, particularly in developing countries. The four genotypes identified in mammals include the G3 found in indigenous hepatitis in countries and regions with high porcine population, and the G1, associated with maternal deaths. Objective: To determine coinfection by hepatitis E virus and the circulating genotypes in Colombia in 1,097 samples using serological markers for hepatitis A, B and C. Materials and methods: Serum samples of 1,097 patients from different regions of Colombia stored at the Laboratorio de Virología of the Instituto Nacional de Salud were selected to detect IgG and IgM anti-hepatitis E virus antibodies. The viral genomes of positive samples were amplified by RT-PCR, and the products were sequenced and phylogenetically analyzed by comparing ORF2 sequences deposited in the GenBank. Results: IgG anti-hepatitis E virus antibodies were found in 278 samples, IgM in 62, and both markers in 64. Hepatitis E virus and hepatitis A virus coinfection determined by IgG anti-hepatitis E virus was 33.6% and 16.1% by IgM; hepatitis E virus and hepatitis B virus coinfection was 23.4% and 8.1%, and hepatitis E virus and hepatitis C virus coinfection was 35.4% and 5.83%, respectively. Among the 52 positive samples by PCR nine were sequenced and grouped within genotype 3A of the American porcine strain. Conclusions: The highest seropositivity was observed for hepatitis A and E. The incidence of hepatitis E virus coinfection with other hepatotropic viruses indicated that this pathogen is more frequent than expected. The circulation of genotype 3A implies that this disease may occur in outbreaks and as zoonosis in Colombia.
Subject(s)
Coinfection , Hepatitis E virus , Genotype , Hepatitis AABSTRACT
Introducción. El virus de la hepatitis A (HAV) es un importante patógeno que se transmite por vía fecal-oral. La epidemiología de la infección está directamente relacionada con el acceso de la población al agua potable y con la infraestructura de alcantarillado. Objetivo. Determinar la presencia del HAV e identificar el genotipo en muestras de agua de abastecimiento y agua residual en ocho municipios, un corregimiento y una vereda del departamento de Antioquia, noroccidente de Colombia. Materiales y métodos. Se hicieron tres muestreos seriados de diciembre de 2012 a abril de 2014 en la fuente principal de abastecimiento de los acueductos y en el principal vertimiento de aguas residuales de cada municipio. Las muestras se concentraron por filtración y ultrafiltración tangencial, y por las técnicas de polietilenglicol y floculación con leche descremada, respectivamente. A partir del ARN total de cada muestra, se amplificaron la región VP3-VP1 para la detección del genoma viral y la región VP1-2B para la genotipificación. Resultados. El genoma del HAV se detectó en las fuentes de agua de abastecimiento de Puerto Berrío, Frontino y Nutibara, y en las muestras de aguas residuales provenientes de los municipios de Arboletes, Zaragoza y Venecia. Mediante el análisis de las secuencias se identificó el subgenotipo IA del virus. Conclusión. Este estudio permitió detectar la presencia del HAV en 6,6 % de las muestras de agua de abastecimiento y en 13,3 % de las muestras de agua residual de los municipios en estudio. Se reporta por primera vez la circulación del subgenotipo IA en muestras ambientales en Antioquia.
Introduction: Hepatitis A virus (HAV) is an important pathogen, typically transmitted via the faecal-oral route. The epidemiology of the infection is directly related to drinking water access and adequate disposal of sewage water. Objective: To determine the presence and identify the genotype of HAV in environmental samples from eight municipalities and two villages in Antioquia, northwestern Colombia. Materials and methods: Three serial samplings were done between December, 2012, and April, 2014. Water samples were obtained from drinking water plants prior to treatment, as well as from the main reserve of wastewater in each municipality included in the study. Viral concentrations for the two types of sample sources were determined by filtration/tangential ultrafiltration and polyethyleneglycol plus flocculation with skimmed milk, respectively. Total ARN was subsequently obtained from each sample and the VP3-VP1 region amplified for detection of the viral genome. The genotype was determined by amplification of the VP1-2B region. Results: The HAV genome was detected in samples from drinking water plants at Puerto Berrío, Frontino and Nutibara, and in wastewater samples from the municipalities of Arboletes, Zaragoza and Venecia. HAV subgenotype IA was identified using phylogenetic analysis. Conclusion: In this study, HAV was identified in 6.6% of the samples from drinking water plants and 13.3% of wastewater samples. This is the first report of HAV subgenotype IA circulating in environmental samples from Antioquia.
Subject(s)
Hepatitis Viruses , Drinking Water , Genotype , Phylogeny , Public Health , WastewaterABSTRACT
Introducción. El agua de consumo humano puede ser vehículo de transmisión de agentes patógenos. La detección de virus entéricos en estas muestras de agua es esencial para establecer las acciones adecuadas de control y prevención de las enfermedades asociadas. Objetivo. Analizar los resultados del diagnóstico de virus entéricos en muestras de agua para el consumo humano recibidas en el Instituto Nacional de Salud y establecer su asociación con los datos sobre la calidad del agua en los municipios de Colombia. Materiales y métodos. Se hizo un análisis descriptivo retrospectivo de los resultados obtenidos en la detección de rotavirus, enterovirus, virus de la hepatitis A y adenovirus, en muestras de agua recibidas para estudios complementarios en la investigación de brotes de hepatitis entérica, de enfermedad diarreica aguda y de enfermedades transmitidas por alimentos. Dicha información se correlacionó con los datos de la vigilancia de la calidad del agua municipal determinada según el índice de riesgo de la calidad del agua (IRCA). Resultados. Se procesaron 288 muestras de 102 municipios de Colombia, de las cuales el 50,7 % fue positivo para algún virus: 26,73 %, para el virus de la hepatitis A; 20,48 %, para enterovirus y rotavirus, y 18,05 % para adenovirus. Se detectaron virus en 48,26 % de las muestras de agua no tratada y en 45,83 % de las de agua tratada. El IRCA no mostró correlación con la presencia de virus. Conclusiones. La presencia de virus en el agua representa un riesgo para la salud pública. La prevención de la transmisión de virus por medio del agua requiere políticas para fortalecer los sistemas de suministro y para mejorar la vigilancia epidemiológica.
Introduction: Since drinking water can be a vehicle for the transmission of pathogens, the detection of enteric viruses in these water samples is essential to establish the appropriate measures to control and prevent associated diseases. Objective: To analyze the results obtained for enteric viruses in water samples for human consumption received at the Colombian Instituto Nacional de Salud and establish their association with the data on water quality in Colombian municipalities. Materials and methods: We conducted a descriptive-retrospective analysis of the results obtained in the detection of rotavirus, enterovirus, hepatitis A virus and adenovirus in water samples received for complementary studies of enteric hepatitis, acute diarrheal disease and foodborne diseases. Data were correlated with the results of water quality surveillance determined by the national human consumption water quality index (IRCA). Results: Of the 288 samples processed from 102 Colombian municipalities, 50.7% were positive for viruses: 26.73% for hepatitis A virus, 20.48% for enterovirus and rotavirus and 18.05% for adenovirus. Viruses were detected in 48.26% of non-treated water samples and in 45.83% of treated water samples. The IRCA index showed no correlation with the presence of viruses. Conclusions: The presence of viruses in water represents a public health risk and, therefore, the prevention of virus transmission through water requires appropriate policies to reinforce water supply systems and improve epidemiological surveillance.
Subject(s)
Enterovirus , Hepatitis A , Rotavirus , Water Pollution , Water SupplyABSTRACT
INTRODUCTION: Hepatitis E virus has emerged as a public health problem, particularly in developing countries. The four genotypes identified in mammals include the G3 found in indigenous hepatitis in countries and regions with high porcine population, and the G1, associated with maternal deaths. OBJECTIVE: To determine coinfection by hepatitis E virus and the circulating genotypes in Colombia in 1,097 samples using serological markers for hepatitis A, B and C. MATERIALS AND METHODS: Serum samples of 1,097 patients from different regions of Colombia stored at the Laboratorio de Virología of the Instituto Nacional de Salud were selected to detect IgG and IgM anti-hepatitis E virus antibodies. The viral genomes of positive samples were amplified by RT-PCR, and the products were sequenced and phylogenetically analyzed by comparing ORF2 sequences deposited in the GenBank. RESULTS: IgG anti-hepatitis E virus antibodies were found in 278 samples, IgM in 62, and both markers in 64. Hepatitis E virus and hepatitis A virus coinfection determined by IgG anti-hepatitis E virus was 33.6% and 16.1% by IgM; hepatitis E virus and hepatitis B virus coinfection was 23.4% and 8.1%, and hepatitis E virus and hepatitis C virus coinfection was 35.4% and 5.83%, respectively. Among the 52 positive samples by PCR nine were sequenced and grouped within genotype 3A of the American porcine strain. CONCLUSIONS: The highest seropositivity was observed for hepatitis A and E. The incidence of hepatitis E virus coinfection with other hepatotropic viruses indicated that this pathogen is more frequent than expected. The circulation of genotype 3A implies that this disease may occur in outbreaks and as zoonosis in Colombia.
Subject(s)
Hepatitis Antibodies/immunology , Hepatitis E virus/immunology , Hepatitis Viruses/immunology , Immunoglobulin G/blood , Immunoglobulin M/blood , Zoonoses/pathology , Animals , Coinfection , Colombia , Genotype , Hepatitis Antibodies/blood , Hepatitis E virus/chemistry , Hepatitis Viruses/chemistry , Humans , Phylogeny , SwineABSTRACT
INTRODUCTION: Hepatitis E virus (HEV) is an emergent virus of global importance; it is the etiological agent of sporadic cases and outbreaks of hepatitis. The epidemiology of this infection in Colombia is unknown. OBJECTIVE: To determine the seropositivity for hepatitis E virus in Colombia in cases with clinical diagnosis of viral hepatitis. MATERIALS AND METHODS: Serum samples from patients that were sent to the Instituto Nacional de Salud during the period 2005-2010 (group 1) and samples sent to the Laboratorio Departamental de Salud Pública de Antioquia during the 2008-2009 period were included in this study (group 2). Serum samples were analyzed by immunoassay with commercial kits. RESULTS: From the 344 analyzed samples, 8.7% were positive for anti-HEV; the frequency of anti-HEV IgM was 1.74% (6/344) and the frequency of anti-HEV IgG was 7.5% (26/344). A difference in frequency of anti-HEV between group 1 (6.3%) and group 2 (1.3%) was observed. The cases were identified in nine departments of Colombia. CONCLUSIONS: This is the first study of hepatitis E virus infection in patients with diagnosis of hepatitis in Colombia. The frequency of anti-HEV described in this population of patients in Colombia is similar to that described in other Latin American countries like Brazil, Perú and Uruguay. Considering the results of this study, it could be necessary to include hepatitis E virus infection serological markers in the differential diagnosis of viral hepatitis in Colombia.
Subject(s)
Hepatitis E/epidemiology , Academies and Institutes , Adolescent , Adult , Child , Child, Preschool , Colombia/epidemiology , Female , Hepatitis Antibodies/blood , Hepatitis E/diagnosis , Hepatitis E virus/immunology , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Laboratories , Male , Middle Aged , Retrospective Studies , Seroepidemiologic Studies , Young AdultABSTRACT
Introducción. El virus de la hepatitis E (HEV), agente etiológico de casos esporádicos y epidemias de hepatitis, es un virus emergente de importancia global. En Colombia se desconoce la epidemiología de la infección causada por este virus. Objetivo. Determinar la seropositividad para el virus de la hepatitis E en muestras de suero de pacientes con diagnóstico clínico de hepatitis viral en Colombia. Materiales y métodos. Se estudiaron muestras de pacientes remitidas al Instituto Nacional de Salud en el periodo 2005-2010 provenientes de 15 departamentos de Colombia (grupo 1) y muestras remitidas al Laboratorio Departamental de Salud Pública de Antioquia en el periodo 2008-2009 (grupo 2). Las muestras de suero se analizaron por inmunoensayo con estuches comerciales. Resultados. La frecuencia de seropositividad para el virus de la hepatitis E en las 344 muestras analizadas fue de 8,7 % (30/344); de estas, 1,74 % (6/344) presentó IgM anti-HEV y 7,5 % (26/344), IgG anti-HEV. Se observó una diferencia en el resultado positivo entre el grupo 1 (6,3 %) y el grupo 2 (15,3 %). Los casos provenían de nueve departamentos del país. Conclusiones. Este es el primer estudio de infección por el virus de la hepatitis E en muestras de pacientes con diagnóstico de hepatitis en Colombia. La seropositividad descrita en esta población de pacientes es similar a la descrita en otros países de América Latina, como Brasil, Perú y Uruguay. Teniendo en cuenta estos resultados, se debe considerar la inclusión de los marcadores de la infección por el virus de la hepatitis E en el diagnóstico diferencial de la hepatitis viral en Colombia.
Introduction: Hepatitis E virus (HEV) is an emergent virus of global importance; it is the etiological agent of sporadic cases and outbreaks of hepatitis. The epidemiology of this infection in Colombia is unknown. Objective: To determine the seropositivity for hepatitis E virus in Colombia in cases with clinical diagnosis of viral hepatitis. Materials and methods: Serum samples from patients that were sent to the Instituto Nacional de Salud during the period 2005-2010 (group 1) and samples sent to the Laboratorio Departamental de Salud Pública de Antioquia during the 2008-2009 period were included in this study (group 2). Serum samples were analyzed by immunoassay with commercial kits. Results: From the 344 analyzed samples, 8.7% were positive for anti-HEV; the frequency of anti-HEV IgM was 1.74% (6/344) and the frequency of anti-HEV IgG was 7.5% (26/344). A difference in frequency of anti-HEV between group 1 (6.3%) and group 2 (1.3%) was observed. The cases were identified in nine departments of Colombia. Conclusions: This is the first study of hepatitis E virus infection in patients with diagnosis of hepatitis in Colombia. The frequency of anti-HEV described in this population of patients in Colombia is similar to that described in other Latin American countries like Brazil, Perú and Uruguay. Considering the results of this study, it could be necessary to include hepatitis E virus infection serological markers in the differential diagnosis of viral hepatitis in Colombia.
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Hepatitis E/epidemiology , Academies and Institutes , Colombia/epidemiology , Hepatitis Antibodies/blood , Hepatitis E virus/immunology , Hepatitis E/diagnosis , Immunoglobulin G/blood , Immunoglobulin M/blood , Laboratories , Retrospective Studies , Seroepidemiologic StudiesABSTRACT
INTRODUCTION: Enteric viruses have been implicated in acute diarrheal disease, food-borne disease, hepatitis A and meningitis outbreaks, in which water was the vehicle of transmission. OBJECTIVE: A concentration method was standardized for the detection of enteric viruses in drinking water. MATERIALS AND METHODS: Twenty liters of water were concentrated to 6 ml by filtration and tangential ultrafiltration. Viral solutions of 20 L each were prepared at 1, 10, 50 and 100 TCID50 of Sabin poliovirus type 1 as positive controls. Viral particles were recovered by tissue culture and detected by conventional polymerase chain reaction (PCR), according to the international standards recommended by the Enterovirus Laboratory at the Centers for Disease Control and Prevention, Atlanta, GA. RESULTS: All positive controls showed cytopathic effect on L20B and RD cells and were amplified by conventional PCR directly from samples. Negative controls did not show any amplification or viral cytopathic effect. CONCLUSIONS: Tangential ultrafiltration for concentrating viruses proved to be a fast, efficient recovery and reproducible. It has the advantage of allowing the detection (at the 1 TCID50 level) and identification of viruses by RT-PCR and the demonstration of viral infectivity by tissue culture.
Subject(s)
Enterovirus/isolation & purification , Water Microbiology/standards , Virology/methodsABSTRACT
Introducción. Los virus entéricos se han visto implicados en brotes de enfermedad diarreica aguda, enfermedades transmitidas por alimentos, hepatitis A y meningitis aséptica, en los que el vehículo de transmisión del agente ha sido el agua.Objetivo. Estandarizar un método de concentración para la detección de virus entéricos en aguas de consumo.Materiales y métodos. Se concentraron 20 litros de agua a un volumen de 6 ml mediante filtración y ultrafiltración tangencial. Como controles positivos se prepararon soluciones de 20 litros a concentraciones virales de 1, 10, 50 y 100 TCID50 (Tissue Culture Infectious Dose 50%) de Poliovirus Sabin de tipo 1. Las partículas virales fueron recuperadas por cultivo en células sensibles a la infección e identificadas por amplificación del genoma viral mediante reacción en cadena de la polimerasa, siguiendo los estándares internacionales de los Centers for Disease Control and Prevention (CDC) de Atlanta. Resultados. Todos los controles positivos causaron efecto citopático en células de rabdomiosarcoma y L20B y fueron detectados por RT- PCR (Real Time- PCR) convencional, directamente de las muestras. Los controles negativos no mostraron efecto citopático ni amplificación viral por RT-PCR.Conclusiones. La ultrafiltración tangencial mostró ser un método rápido y eficaz al recuperar virus desde una TCID50, además de ser reproducible y sencillo. Tiene la ventaja de permitir la detección de su capacidad de contagiosidad viral por el cultivo celular, y la identificación por RT-PCR.
Introduction. Enteric viruses have been implicated in acute diarrheal disease, food-borne disease, hepatitis A and meningitis outbreaks, in which water was the vehicle of transmission.Objective. A concentration method was standardized for the detection of enteric viruses in drinking water. Materials and methods. Twenty liters of water were concentrated to 6 ml by filtration and tangential ultrafiltration. Viral solutions of 20 L each were prepared at 1, 10, 50 and 100 TCID50 of Sabin poliovirus type 1 as positive controls. Viral particles were recovered by tissue culture and detected by conventional polymerase chain reaction (PCR), according to the international standards recommended by the Enterovirus Laboratory at the Centers for Disease Control and Prevention, Atlanta, GA. Results. All positive controls showed cytopathic effect on L20B and RD cells and were amplified by conventional PCR directly from samples. Negative controls did not show any amplification or viral cytopathic effect. Conclusions. Tangential ultrafiltration for concentrating viruses proved to be a fast, efficient recovery and reproducible. It has the advantage of allowing the detection (at the 1 TCID50 level) and identification of viruses by RT-PCR and the demonstration of viral infectivity by tissue culture.
Subject(s)
Cytopathogenic Effect, Viral , Enterovirus , Poliovirus , Reverse Transcriptase Polymerase Chain Reaction , Water QualityABSTRACT
OBJECTIVES: To establish an in-hospital surveillance system for diarrhea in children under five, to estimate the burden of rotavirus-related disease, and to identify the most common rotavirus genotypes. METHODS: Included in the study were children who were hospitalized for serious complications of diarrhea in three medical care facilities in Bogotá, Barranquilla, and Cali, Colombia. A solid-phase enzyme-linked immunosorbent assay (ELISA) was used to detect rotavirus, and reverse-transcriptase polymerase chain reaction (RT-PCR) was the genotyping method employed. The frequencies, central tendency, and dispersion of the variables were determined. Stratified analysis and bivariate analysis were performed by applying a chi squared test or Fisher's exact test, or a chi squared test for trends, depending on the type of data analyzed. Relative risks were established. For analyzing trends we performed linear regression and calculated correlation coefficients and P values. RESULTS: Between December 2003 and November 2004, 893 children were hospitalized in the three participating centers included in the study. Of these children, 68% were between 6 and 23 months of age; 2.7% of hospitalized patients showed clinical signs of hypovolemic shock, and 1.2% died. Only 57% of the mothers had given their children an oral rehydration solution before hospitalization. Rotavirus infection was the cause of 50% of hospitalizations (correlation coefficient [r] > 0.8) and was linked to an inability to hold down orally-ingested fluids (relative risk [RR] = 1.45; 95% confidence interval [95% CI]: 1.24 to 1.69; P < 0.0000) and to intractable vomiting (RR = 1.47; 95% CI: 1.16 to 1.86; P < 0.01). Rotavirus infection led to one death per 2 000 children; 16 hospitalizations per 1 000 children, and 631 medical visits per 1 000 children. A seasonal trend was noted for G1P[8], G2P[4], and G3P[8] rotavirus genotypes, and this did not vary as a result of geographic distance or differences in temperature, humidity, and rainfall among cities. CONCLUSIONS: Rotavirus infection is an important cause of morbidity and mortality from diarrhea, particularly during the first years of life, when children are more susceptible to serious complications. It is important for prevention strategies to have a high impact before 6 months of age, and vaccination against rotavirus can be a good complementary intervention strategy. No description was found in the international scientific literature of the seasonal variations in rotavirus genotypes. It is important to carry out cost-effectiveness studies in order to promote the investment of resources in accordance with population needs, and to continue surveillance activities so as to better understand how the virus behaves.
Subject(s)
Diarrhea, Infantile/epidemiology , Diarrhea, Infantile/virology , Rotavirus Infections/epidemiology , Child, Preschool , Colombia , Diarrhea, Infantile/therapy , Female , Hospitalization , Humans , Infant , Male , Rotavirus/isolation & purification , Rotavirus Infections/complications , Rotavirus Infections/therapy , Rotavirus Infections/virologyABSTRACT
OBJETIVOS: Establecer un sistema de vigilancia intrahospitalaria de la diarrea en niños menores de 5 años, estimar la carga de la enfermedad por rotavirus e identificar los genotipos más frecuentes de rotavirus. MÉTODOS: Se incluyó en este estudio a niños hospitalizados por complicaciones graves de la diarrea en tres centros asistenciales de Santa Fe de Bogotá, Barranquilla y Cali, Colombia. Se utilizó un método de enzimoinmunoanálisis en fase sólida (ELISA) para la detección de rotavirus y un método de reacción en cadena de la polimerasa con transcripción inversa (RT-PCR) para la genotipificación. Se determinaron las frecuencias, la tendencia central y la dispersión de las variables. Se realizó un análisis estratificado y un análisis con dos variables, mediante una prueba de la ji2 o una prueba exacta de Fisher, o una prueba de la ji2 para evaluar la tendencia, según los datos. Se establecieron los riesgos relativos. Para el análisis de la tendencia, se utilizaron la regresión lineal, los coeficientes de correlación y los valores de P. RESULTADOS: Entre diciembre de 2003 y noviembre de 2004 se hospitalizó a 893 niños en los tres centros participantes en el estudio, de los cuales el 68 por ciento tenía entre 6 y 23 meses de edad. Un 2,7 por ciento de los pacientes ingresados presentaba un cuadro de choque hipovolémico y un 1,2 por ciento falleció. Solo un 57 por ciento de las madres había administrado a sus hijos una solución de rehidratación oral antes de la hospitalización. La infección por rotavirus motivó un 50 por ciento de las hospitalizaciones (coeficiente de correlación [r] > 0,8) y se relacionó con intolerancia a la vía oral (riesgo relativo [RR] = 1,45; intervalo de confianza del 95 por ciento [IC95 por ciento]: 1,24 - 1,69; P < 0,0000) y vómito incoercible (RR = 1,47; IC95 por ciento: 1,16 - 1,86; P < 0,01). La infección por rotavirus ocasionó una muerte por cada 2 000 niños; 16 hospitalizaciones por cada 1 000 niños y 631 consultas por cada 1 000 niños. Se identificó estacionalidad en los genotipos G1P[8], G2P[4] y G3P[8] de rotavirus, que no varió a pesar de la distancia geográfica y las diferencias de temperatura, humedad y precipitación entre las tres ciudades.CONCLUSIONES: La infección por rotavirus es una causa importante de morbilidad y mortalidad por diarrea, especialmente en los primeros años de vida, cuando los niños están más expuestos a las complicaciones graves. Es necesario que las estrategias de prevención tengan un alto impacto antes de los 6 meses de edad. La vacunación contra el rotavirus puede ser una buena estrategia complementaria de intervención. No se encontró en la literatura internacional ninguna descripción anterior de la estacionalidad de los genotipos de rotavirus. Es importante hacer estudios de costo-efectividad para favorecer la inversión de recursos según las necesidades de la población y continuar la vigilancia para ampliar el conocimiento del comportamiento de este virus.
OBJECTIVES: To establish an in-hospital surveillance system for diarrhea in children under five, to estimate the burden of rotavirus-related disease, and to identify the most common rotavirus genotypes. METHODS: Included in the study were children who were hospitalized for serious complications of diarrhea in three medical care facilities in Bogotá, Barranquilla, and Cali, Colombia. A solid-phase enzyme-linked immunosorbent assay (ELISA) was used to detect rotavirus, and reverse-transcriptase polymerase chain reaction (RT-PCR) was the genotyping method employed. The frequencies, central tendency, and dispersion of the variables were determined. Stratified analysis and bivariate analysis were performed by applying a chi squared test or Fisher's exact test, or a chi squared test for trends, depending on the type of data analyzed. Relative risks were established. For analyzing trends we performed linear regression and calculated correlation coefficients and P values. RESULTS: Between December 2003 and November 2004, 893 children were hospitalized in the three participating centers included in the study. Of these children, 68 percent were between 6 and 23 months of age; 2.7 percent of hospitalized patients showed clinical signs of hypovolemic shock, and 1.2 percent died. Only 57 percent of the mothers had given their children an oral rehydration solution before hospitalization. Rotavirus infection was the cause of 50 percent of hospitalizations (correlation coefficient [r] > 0.8) and was linked to an inability to hold down orally-ingested fluids (relative risk [RR] = 1.45; 95 percent confidence interval [95 percent CI]: 1.24 to 1.69; P < 0.0000) and to intractable vomiting (RR = 1.47; 95 percent CI: 1.16 to 1.86; P < 0.01). Rotavirus infection led to one death per 2 000 children; 16 hospitalizations per 1 000 children, and 631 medical visits per 1 000 children. A seasonal trend was noted for G1P[8], G2P[4], and G3P[8] rotavirus genotypes, and this did not vary as a result of geographic distance or differences in temperature, humidity, and rainfall among cities.CONCLUSIONS: Rotavirus infection is an important cause of morbidity and mortality from diarrhea, particularly during the first years of life, when children are more susceptible to serious complications. It is important for prevention strategies to have a high impact before 6 months of age, and vaccination against rotavirus can be a good complementary intervention strategy. No description was found in the international scientific literature of the seasonal variations in rotavirus genotypes. It is important to carry out cost-effectiveness studies in order to promote the investment of resources in accordance with population needs, and to continue surveillance activities so as to better understand how the virus behaves.
Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Diarrhea, Infantile/epidemiology , Diarrhea, Infantile/virology , Rotavirus Infections/epidemiology , Colombia , Diarrhea, Infantile/therapy , Hospitalization , Rotavirus Infections/complications , Rotavirus Infections/therapy , Rotavirus Infections/virology , Rotavirus/isolation & purificationABSTRACT
Objetivo. Realizar el seguimiento de la infección in vitro por enterovirus 71 (EV71) sobre celulas TE-671, evaluando la presencia de los antígenos virales y la pérdida de viabilidad en diferentes tiempos de infección. Materiales y métodos. Se infectaron cultivos de células TE-671 con diferentes diluciones de EV71, aislado de un caso de parálisis flácida aguda (3364-COL-94), y por varios tiempos posinfección (6,12,24,48,72 horas). Los cultivos se procesaron por una técnica de inmunoperoxidasa indirecta para obtener las proporciones de infección en cada condición. Simultáneamente, se realizó una prueba de viabilidad por el método bioquímico del MTT para evaluar la pérdida de la viabilidad celular debida a la infección. Resultados. Aunque por inmunocitoquímica se detectó infección desde las 6 horas después de la inoculación (pi), el efecto citopático y la mortalidad celular fueron drásticos después de 48 horas pi. De esta manera, el mejor tiempo para la detección de los antígenos virales fue a las 12 horas pi, momento en el cual la muerte celular no es tan grave, lo cual permite una adecuada detección del antígeno viral. Conclusiones. Se ratificó la alta receptividad de las células TE-671 a la infección por EV71, la cual se correlaciona con el importante efecto citopático evidenciando por la prueba de MTT, lo cual convierte a esta línea celular e un recurso eficiente para el aislamiento y la identificación de EV71 y, también, para estudios de su biología básica
Subject(s)
Enterovirus , In Vitro Techniques , Enterovirus Infections , T-Lymphocytes , Nerve Growth Factors , Cell Culture TechniquesABSTRACT
OBJECTIVES: To establish the relationship between rotavirus infection and dehydration from diarrhea in Colombian children under 5 years of age, and to identify risk factors for diarrhea with dehydration. METHODS: A case-control study was performed in an urban hospital in Bogota, Colombia, between April 2000 and February 2001. The sample was composed of 290 children of both sexes under 5 years of age; of these children, 145 of them were hospitalized for acute diarrheal disease (ADD) with dehydration (cases), and 145 had a diagnosis of ADD but no signs of dehydration (controls). All children underwent a complete physical examination. Mothers responded to a questionnaire containing items on demographic and socioeconomic variables, as well as on knowledge, attitudes, and practices with regard to hygiene, and on access to health services. RESULTS: An association was detected between diarrhea with dehydration and the presence of rotavirus in fecal samples (odds ratio [OR] = 3.46; 95% confidence interval [95% CI]: 1.71 to 7.00), birth weight < 2 600 g (OR = 7.79; 95% CI: 3.47 to 18.01), and breastfeeding for less than 3 months (OR = 3.17; 95% CI: 1.66 to 6.13). The risk of having dehydration was associated with low socioeconomic status, poor hygienic practices among the child's family members, and mother's low educational level. CONCLUSIONS: The ineffectiveness of health promotion and disease prevention activities in a population with easy access to health services set the stage for the appearance of cases of diarrhea with dehydration. Rotavirus infection plays an important role in the severity of ADD among Colombian children.
Subject(s)
Dehydration/virology , Diarrhea, Infantile/virology , Rotavirus Infections/complications , Rotavirus/isolation & purification , Acute Disease , Case-Control Studies , Child, Preschool , Colombia/epidemiology , Dehydration/epidemiology , Diarrhea, Infantile/epidemiology , Female , Humans , Infant , Male , Mothers , Public Health , Risk Factors , Rotavirus Infections/epidemiology , Socioeconomic FactorsABSTRACT
Objetivos. Establecer la relación entre la infección por rotavirus y la deshidratación por diarrea en niños colombianos menores de 5 años y determinar los factores de riesgo de la diarrea con deshidratación. Métodos. Se realizó un estudio de casos y testigos en un hospital urbano de Bogotá, Colombia, entre abril de 2000 y febrero de 2001. Se seleccionaron 290 niños menores de 5 años de edad de uno u otro sexo; de ellos 145 estaban hospitalizados por enfermedad diarreica aguda (EDA) con deshidratación (casos) y 145 tenían diagnóstico de EDA sin signos de deshidratación (testigos). A todos los niños se les realizó un examen físico completo. Las madres respondieron un cuestionario con variables demográficas; socioeconómicas; de conocimientos, actitudes y hábitos higiénicos; y de acceso a los servicios de salud. Se determinó la presencia de rotavirus en muestras de heces fecales tomadas durante las primeras 48 horas del ingreso al estudio. Resultados. Se encontró una asociación estadística entre la diarrea con deshidratación y la presencia de rotavirus en las muestras de heces (razón de posibilidades [RP] = 3,46; intervalo de confianza de 95% [IC95%]: 1,71 a 7,00), el peso al nacer < 2 600 g (RP = 7,79; IC95%: 3,47 a 18,01) y el tiempo de lactancia materna inferior a 3 meses (RP = 3,17; IC95%: 1,66 a 6,13). El riesgo de presentar deshidratación se asoció con las condiciones socioeconómicas desfavorables, los hábitos higiénicos inadecuados de la familia del niño y una menor escolaridad materna. Conclusiones. La calidad inadecuada de las acciones de promoción y prevención en una población con fácil acceso a los servicios de salud favoreció la aparición de casos de diarrea con deshidratación. La infección por rotavirus desempeña un papel importante en la gravedad de la EDA en niños colombianos
Objectives. To establish the relationship between rotavirus infection and dehydration from diarrhea in Colombian children under 5 years of age, and to identify risk factors for diarrhea with dehydration. Methods. A case-control study was performed in an urban hospital in Bogotá, Colombia, between April 2000 and February 2001. The sample was composed of 290 children of both sexes under 5 years of age; of these children, 145 of them were hospitalized for acute diarrheal disease (ADD) with dehydration (cases), and 145 had a diagnosis of ADD but no signs of dehydration (controls). All children underwent a complete physical examination. Mothers responded to a questionnaire containing items on demographic and socioeconomic variables, as well as on knowledge, attitudes, and practices with regard to hygiene, and on access to health services. Results. An association was detected between diarrhea with dehydration and the presence of rotavirus in fecal samples (odds ratio [OR] = 3.46; 95% confidence interval [95% CI]: 1.71 to 7.00), birth weight < 2 600 g (OR = 7.79; 95% CI: 3.47 to 18.01), and breastfeeding for less than 3 months (OR = 3.17; 95% CI: 1.66 to 6.13). The risk of hav ing dehydration was associated with low socioeconomic status, poor hygienic practices among the child's family members, and mother's low educational level. Conclusions. The ineffectiveness of health promotion and disease prevention activ ities in a population with easy access to health services set the stage for the appearance of cases of diarrhea with dehydration. Rotavirus infection plays an important role in the severity of ADD among Colombian children
Subject(s)
Risk Factors , Rotavirus , Diarrhea, Infantile , Colombia , DehydrationABSTRACT
Las parálisis fláccidas agudas (PFA) tienen una amplia variedad de orígenes y de agentes causales: físicos, fisiopatológicos, tóxicos e infecciosos. Entre estos últimos, el virus salvaje (silvestre) de la poliomielitis y el enterovirus 71(EV71), parecen ser los agentes virales más frecuentes. Habiendo eliminado el poliovirus salvaje autóctono como agente causal de enfermedad paralítica en Colombia desde junio de 1991 y teniendo aislamientos de virus no polio en el 20,8 por ciento del total de casos de PFA notificados anualmente, quisimos conocer el papel que juegan los enterovirus en la incidencia de parálisis fláccida aguda y la dinámica de circulación y distribución de los mismos en Colombia. Se revisó la base de datos de los casos notificados al Programa de Erradicación de la Poliomielitis en Colombia entre el 1º de enero de 1992 y el 31 de diciembre de 1995, al cual se notificaron 856 casos sospechosos de niños menores de 15 años, y se escogieron 69 casos para el estudio pero se recuperaron 57 aislamientos virales por reinoculación en células RD y Hep-2C. Estos se identificaron mediante neutralización con mezclas de antisueros de Lim & Benyesh-Melnick (LBM). Todos ellos fueron sometidos a caracterización molecular mediante reacción en cadena de la polimerasa -PCR-, utilizando iniciadores complementarios a la region VP1 del genoma viral, seguida del análisis de secuencia de nucleótidos de los fragmentos amplificados por PCR. La identificación final del serotipo se realizó por comparación de nucleótidos en auto assembler y el análisis descriptivo de los datos. No se estableció circulación mayor de ningún serotipo específico en región geográfica alguna del país. Tampoco hubo asociación causal a ninguna patología característica con ninguno de los enterovirus aislados. Se describe el hallazgo de EV71 en un caso de PFA con diagnóstico clínico de síndrome de Guillain-Barre. La descripción de 22 serotipos diferentes de enterovirus no polio que circulan en Colombia (19 serotipos identificados por métodos moleculares y 3 por seroneutralización), coincide con los serotipos más frecuentemente descritos en otros estudios. Los serotipos Coxsackievirus B5, B1, B3, Echovirus 6, 7, 13, 20, 30, Coxsackievirus A2, A10, A14, A16, A18, A21, fueron los serotipos de enterovirus más frecuentes en Colombia durante el período 1992-1995