RESUMO
Torque Teno Virus (TTV) is a nonpathogenic and ubiquitous ssDNA virus, a member of the Anelloviridae family. TTV has been postulated as a biomarker in transplant patients. This study aimed to determine the TTV species diversity and variability in renal transplant recipients and to associate species diversity with the corresponding TTV viral load. From 27 recipients, 30 plasma samples were selected. Viral load was determined using two real-time PCR assays, followed by RCA-NGS and ORF1 phylogenetic analysis. The TTV diversity was determined in all samples. Variability was determined in three patients with two sequential samples (pre- and post-transplantation). Most of the samples presented multiple TTV species, up to 15 different species were detected. In the pre-transplant samples (n = 12), the most prevalent species were TTV3 (75%) and TTV13 (75%), and the median number of species per sample was 5 (IQR: 4-7.5). TTV3 was also the most prevalent (56%) in the post-transplant samples (n = 18), and the median number of species was 2 (IQR: 1.8-5.5). No significant correlation between the number of species and viral load was found. The number and type of TTV species showed total variability over time. We report high TTV species diversity in Argentinian recipients, especially in pre-transplant period, with total intra-host variability. However, we found no significant correlation between this high diversity and TTV viral load.
Assuntos
Infecções por Vírus de DNA , Transplante de Rim , Torque teno virus , Humanos , Torque teno virus/genética , Transplante de Rim/efeitos adversos , Filogenia , Transplantados , Carga Viral , DNA Viral/genéticaRESUMO
Abstract Asymptomatic infections with SARS-CoV-2 are associ ated with viral transmission and have a key role in the propagation of the pandemic. Understanding viral shed ding during asymptomatic infections is critical. Unfor tunately, data on asymptomatic SARS-CoV-2 infection in children is extremely limited. To determine the presence of viral viable shedding, we prospectively followed two healthy children of a family where both parents devel oped mild COVID-19 (April 2021). SARS-CoV-2 detection was made by RT-PCR and virus isolation by cell culture from saliva samples. Positive samples were sequenced to identify variants of SARS-CoV-2. Serum samples were evaluated to determine the presence of antibodies using a single enzyme-linked immunosorbent assay (ELISA, COVIDAR IgG). Both children were SARS-CoV-2 positive and asymptomatic. In addition, the virus grew in cell cul ture from saliva samples. Furthermore, one child showed viable SARS-CoV-2 for at least 17 days after the onset symptoms from his father. The recommended isolation period for asymptomatic contacts during the acquisition of data had been established for 10 days; however, this child remained with viable virus beyond that period. The positive samples from both children were consistent with B.1.1.28.1 lineage (Gamma). In both asymptomatic children, anti-Spike IgG was detected. Asymptomatic children may represent a source of infection that should not be underestimated during this pandemic.
Resumen Las infecciones asintomáticas por SARS-CoV-2 están asociadas a la transmisión viral y tienen un papel cla ve en la propagación de la pandemia. Comprender la excreción viral durante las infecciones asintomáticas es fundamental. Desafortunadamente, los datos sobre la infección asintomática por SARS-CoV-2 en niños son extremadamente limitados. Para determinar la presencia de excreción de virus viable, se siguió prospectivamente a dos niños sanos de una familia en la que ambos padres desarrollaron COVID-19 leve (abril 2021). La detección de SARS-CoV-2 se realizó por RT-PCR y el aislamiento del virus por cultivo celular a partir de muestras de saliva. Las muestras positivas se secuenciaron para identificar variantes de SARS-CoV-2. En las muestras de suero se determinó la presencia de anticuerpos utilizando un ensayo de ELISA (COVIDAR IgG). Ambos niños fueron positivos para SARS-CoV-2 y asintomáticos. Además, el virus creció en cultivos celulares a partir de muestras de saliva. Uno de los niños mantuvo SARS-CoV-2 via bles durante al menos 17 días después de la aparición de los síntomas de su padre. El período de aislamiento recomendado para contactos asintomáticos durante la adquisición de datos se había establecido en 10 días, sin embargo, este niño permaneció con virus viable más allá de ese período. Las muestras positivas de estos niños correspondieron al linaje B.1.1.28.1 (Gamma). En ambos niños asintomáticos se detectó anticuerpos IgG anti-Spike. Concluimos que los niños asintomáticos pueden representar una fuente de infección que no debe subestimarse durante esta pandemia.
RESUMO
Asymptomatic infections with SARS-CoV-2 are associated with viral transmission and have a key role in the propagation of the pandemic. Understanding viral shedding during asymptomatic infections is critical. Unfortunately, data on asymptomatic SARS-CoV-2 infection in children is extremely limited. To determine the presence of viral viable shedding, we prospectively followed two healthy children of a family where both parents developed mild COVID-19 (April 2021). SARS-CoV-2 detection was made by RT-PCR and virus isolation by cell culture from saliva samples. Positive samples were sequenced to identify variants of SARS-CoV-2. Serum samples were evaluated to determine the presence of antibodies using a single enzyme-linked immunosorbent assay (ELISA, COVIDAR IgG). Both children were SARS-CoV-2 positive and asymptomatic. In addition, the virus grew in cell culture from saliva samples. Furthermore, one child showed viable SARS-CoV-2 for at least 17 days after the onset symptoms from his father. The recommended isolation period for asymptomatic contacts during the acquisition of data had been established for 10 days; however, this child remained with viable virus beyond that period. The positive samples from both children were consistent with B.1.1.28.1 lineage (Gamma). In both asymptomatic children, anti-Spike IgG was detected. Asymptomatic children may represent a source of infection that should not be underestimated during this pandemic.
Las infecciones asintomáticas por SARS-CoV-2 están asociadas a la transmisión viral y tienen un papel clave en la propagación de la pandemia. Comprender la excreción viral durante las infecciones asintomáticas es fundamental. Desafortunadamente, los datos sobre la infección asintomática por SARS-CoV-2 en niños son extremadamente limitados. Para determinar la presencia de excreción de virus viable, se siguió prospectivamente a dos niños sanos de una familia en la que ambos padres desarrollaron COVID-19 leve (abril 2021). La detección de SARS-CoV-2 se realizó por RT-PCR y el aislamiento del virus por cultivo celular a partir de muestras de saliva. Las muestras positivas se secuenciaron para identificar variantes de SARS-CoV-2. En las muestras de suero se determinó la presencia de anticuerpos utilizando un ensayo de ELISA (COVIDAR IgG). Ambos niños fueron positivos para SARS-CoV-2 y asintomáticos. Además, el virus creció en cultivos celulares a partir de muestras de saliva. Uno de los niños mantuvo SARS-CoV-2 viables durante al menos 17 días después de la aparición de los síntomas de su padre. El período de aislamiento recomendado para contactos asintomáticos durante la adquisición de datos se había establecido en 10 días, sin embargo, este niño permaneció con virus viable más allá de ese período. Las muestras positivas de estos niños correspondieron al linaje B.1.1.28.1 (Gamma). En ambos niños asintomáticos se detectó anticuerpos IgG anti-Spike. Concluimos que los niños asintomáticos pueden representar una fuente de infección que no debe subestimarse durante esta pandemia.
Assuntos
COVID-19 , SARS-CoV-2 , Criança , Humanos , Infecções Assintomáticas , Anticorpos Antivirais , Imunoglobulina GRESUMO
Abstract Health care workers (HCWs) are at high risk for SARS-CoV-2. In addition, pre-symptomatic or asymptomatic transmission accounts for around half of the cases. Saliva testingis an option to detect SARS-CoV-2 infection. To determine the performance of saliva samplesfor screening, HCWs were tested for SARS-CoV-2 by RT-PCR. Those with a positive result insaliva were tested by nasopharyngeal swabbing for viral RNA detection and blood collectionto search for the presence of specific antibodies. In September---October 2020, 100 HCWs wereenrolled and followed up. Six subjects (6%) tested positive in saliva. Of them, 5/6 were positivein a subsequent nasopharyngeal swab and 4/6 developed signs and symptoms compatible withCOVID-19. Among the latter, 3 seroconverted while asymptomatic HCWs remained seronega-tive. Saliva screening was helpful for identifying SARS-CoV-2 infection in HCWs. This screeningpermitted rapid personnel isolation avoiding further transmission of the virus in the hospitalsetting.
Resumen El personal de salud (PS) tiene un alto riesgo de contraer SARS-CoV-2. La transmisión presintomática/asintomática representa alrededor de la mitad de los casos y el análisis a partir de muestras de saliva puede ser una opción para detectar la infección. Para determinar el rendimiento de estas muestras, 100 voluntarios del PS se sometieron a la detección de SARS-CoV-2 por RT-PCR en muestras de saliva en el período septiembre-octubre de 2020. De aquellos con resultado positivo en saliva, se tomaron hisopados nasofaríngeos para detectar ARN viral y muestras de suero para evaluar anticuerpos específicos. Se detectó ARN viral en la saliva de seis individuos (6%). De ellos, 5/6 fueron SARS-CoV-2 positivos en hisopado nasofaríngeo y 4/6 desarrollaron signos y síntomas compatibles con COVID-19. Entre estos últimos, tres serocon-virtieron, en tanto que los voluntarios asintomáticos permanecieron seronegativos. La muestra de saliva fue útil para identificar la infección por SARS-CoV-2 en esta cohorte del personal de salud y así proceder al rápido aislamiento de los individuos infectados, lo que evitó una mayor transmisión del virus en el ámbito hospitalario.
RESUMO
Health care workers (HCWs) are at high risk for SARS-CoV-2. In addition, pre-symptomatic or asymptomatic transmission accounts for around half of the cases. Saliva testing is an option to detect SARS-CoV-2 infection. To determine the performance of saliva samples for screening, HCWs were tested for SARS-CoV-2 by RT-PCR. Those with a positive result in saliva were tested by nasopharyngeal swabbing for viral RNA detection and blood collection to search for the presence of specific antibodies. In September-October 2020, 100 HCWs were enrolled and followed up. Six subjects (6%) tested positive in saliva. Of them, 5/6 were positive in a subsequent nasopharyngeal swab and 4/6 developed signs and symptoms compatible with COVID-19. Among the latter, 3 seroconverted while asymptomatic HCWs remained seronegative. Saliva screening was helpful for identifying SARS-CoV-2 infection in HCWs. This screening permitted rapid personnel isolation avoiding further transmission of the virus in the hospital setting.
Assuntos
COVID-19 , SARS-CoV-2 , Humanos , COVID-19/diagnóstico , Saliva , Pessoal de Saúde , NasofaringeRESUMO
Human adenoviruses (HAdV) are one of the most frequent causes of respiratory infections around the world, causing mild to severe disease. In Argentina, many studies focused on the association of HAdV respiratory infection with severe disease and fatal outcomes leading to the discovery in 1984 of a genomic variant 7h associated with high fatality. Although several molecular studies reported the presence of at least 4 HAdV species (B, C, D and E) in Argentina, few sequences were available in the databases. In this study, sequences from the hexon gene region were obtained from 141 patients as a first approach to assess the genetic diversity of HAdVs circulating in Buenos Aires, Argentina. Phylogenetic analysis of these sequences and others recovered from public databases confirmed the circulation of the four above-mentioned species represented by 11 genotypes, with predominance in species B and C and shifts in their proportion in the studied period (2000 to 2018). The variants detected in Argentina, for most of the genotypes, were similar to those already described in other countries. However, uncommon lineages belonging to genotypes C2, C5 and E4 were detected, which might indicate the circulation of local variants and will deserve further studies of whole-genome sequences.
Assuntos
Infecções por Adenovirus Humanos/genética , Adenovírus Humanos/genética , Bases de Dados de Ácidos Nucleicos , Genótipo , Filogenia , Infecções Respiratórias/genética , Infecções por Adenovirus Humanos/epidemiologia , Adenovírus Humanos/isolamento & purificação , Argentina/epidemiologia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Análise de Sequência de DNARESUMO
Current diagnostic standards involve severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection in nasopharyngeal swabs (NPS), but saliva is an attractive and noninvasive option for diagnosis. The objectives were to determine the performance of saliva in comparison with NPS for detecting SARS-CoV-2 and to compare the optimized home brew reverse-transcription polymerase chain reaction (RT-PCR) with a commercial RT-PCR. Paired NPS and saliva specimens were prospectively collected and tested by RT-PCR from patients presenting at an emergency room with signs and symptoms compatible with coronavirus disease-2019. A total of 348 samples from 174 patients were tested by RT-PCR assays. Among 174 patients with symptoms, 63 (36%) were SARS-CoV-2 positive in NPS using the optimized home-brew PCR. Of these 63 patients, 61 (98%) were also positive in saliva. An additional positive SARS-CoV-2 saliva was detected in a patient with pneumonia. Kappa Cohen's coefficient agreement between NPS and saliva was 0.96 (95% confidence interval [CI], 0.90-0.99). Median Ct values in NPS versus saliva were 18.88 (interquartile range [IQR], 15.60-23.58; range, 11.97-38.10) versus 26.10 (IQR, 22.75-30.06; range, 13.78-39.22), respectively (p < .0001). The optimized home-brew RT-PCR demonstrated higher analytical and clinical sensitivity compared with the commercial RT-PCR assay. A high sensitivity (98%) and agreement (kappa 0.96) in saliva samples compared to NPS was demonstrated when using an optimized home-brew PCR even when the viral load in saliva was lower than in NPS. This noninvasive sample is easy to collect, requires less consumable and avoids discomfort to patients. Importantly, self-collection of saliva can diminish exposure to healthcare personnel.
Assuntos
COVID-19/diagnóstico , COVID-19/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , SARS-CoV-2/isolamento & purificação , Saliva/virologia , Manejo de Espécimes/métodos , Adulto , Idoso , Serviço Hospitalar de Emergência , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
We report the emergency development and application of a robust serologic test to evaluate acute and convalescent antibody responses to SARS-CoV-2 in Argentina. The assays, COVIDAR IgG and IgM, which were produced and provided for free to health authorities, private and public health institutions and nursing homes, use a combination of a trimer stabilized spike protein and the receptor binding domain (RBD) in a single enzyme-linked immunosorbent assay (ELISA) plate. Over half million tests have already been distributed to detect and quantify antibodies for multiple purposes, including assessment of immune responses in hospitalized patients and large seroprevalence studies in neighborhoods, slums and health care workers, which resulted in a powerful tool for asymptomatic detection and policy making in the country. Analysis of antibody levels and longitudinal studies of symptomatic and asymptomatic SARS-CoV-2 infections in over one thousand patient samples provided insightful information about IgM and IgG seroconversion time and kinetics, and IgM waning profiles. At least 35% of patients showed seroconversion within 7 days, and 95% within 45 days of symptoms onset, with simultaneous or close sequential IgM and IgG detection. Longitudinal studies of asymptomatic cases showed a wide range of antibody responses with median levels below those observed in symptomatic patients. Regarding convalescent plasma applications, a protocol was standardized for the assessment of end point IgG antibody titers with COVIDAR with more than 500 plasma donors. The protocol showed a positive correlation with neutralizing antibody titers, and was used for clinical trials and therapies across the country. Using this protocol, about 80% of convalescent donor plasmas were potentially suitable for therapies. Here, we demonstrate the importance of providing a robust and specific serologic assay for generating new information about antibody kinetics in infected individuals and mitigation policies to cope with pandemic needs.
Assuntos
COVID-19/virologia , SARS-CoV-2/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Adulto , Idoso , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Formação de Anticorpos , Argentina/epidemiologia , COVID-19/epidemiologia , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Pandemias , SARS-CoV-2/isolamento & purificação , Estudos SoroepidemiológicosRESUMO
OBJECTIVE: To report a conjunctivitis outbreak in a neonatology intensive care unit (NICU) and determine the associated economic impact. DESIGN: Prospective observational study. SETTING: Centro de Educación Médica e Investigaciones Clínicas (CEMIC) University Hospital, a private, tertiary-care healthcare institution in Buenos Aires, Argentina. PARTICIPANTS: The study included 52 NICU neonates and 59 NICU-related healthcare workers (HCWs) from CEMIC hospital. METHODS: Neonates and HCWs were swabbed for real-time polymerase chain reaction (PCR) testing, viral culture, and typing by sequencing. Infection control measures, structural and logistic changes were implemented. Billing records were analyzed to determine costs. RESULTS: From January 30 to April 28, 2018, 52 neonates were hospitalized in the NICU. Among them, 14 of 52 (21%) had bilateral conjunctivitis with pseudomembranes. Symptomatic neonates and HCWs were HAdV-D8 positive. Ophthalmological symptoms had a median duration of 18 days (IQR, 13-24.5). PCR positivity and infectious range had a median duration of 18.5 days. As part of containment measures, the NICU and the high-risk pregnancy unit were closed to new patients. The NICU was divided into 2 areas for symptomatic and asymptomatic patients; a new room was assigned for the general nursery, and all deliveries from the high-risk pregnancy unit were redirected to other hospitals. The outbreak cost the hospital US$205,000: implementation of a new nursery room and extra salaries cost US$30,350 and estimated productivity loss during 1 month cost US$175,000. CONCLUSIONS: Laboratory diagnosis confirmed the cause of this outbreak as HAdV-D8. The immediate adoption and reinforcement of rigorous infection control measures limited the nosocomial viral spread. This outbreak represented a serious institutional problem, causing morbidity, significant economic loss, and absenteeism.
Assuntos
Conjuntivite , Infecção Hospitalar , Neonatologia , Adenoviridae , Conjuntivite/epidemiologia , Infecção Hospitalar/epidemiologia , Surtos de Doenças , Feminino , Genótipo , Humanos , Recém-Nascido , Controle de Infecções , Unidades de Terapia Intensiva Neonatal , GravidezRESUMO
Lower acute respiratory infections (ARI) are a frequent cause of morbidity and mortality in infants, respiratory viruses being the major causative agents. The aim of this work was to determine the respiratory pathogen frequency, the clinical characteristics and the outcome in infants <2 months old hospitalized with ARI. A retrospective study was performed during a five-year period (2008-2011, 2014-2016). Respiratory viruses and atypical bacteria were studied using the FilmArray-Respiratory Panel. Demographic and clinical characteristics, hospitalization course and outcomes were evaluated. Of the 137 infants <2 months old hospitalized with ARI studied, a 94.9% positivity rate as determined in 117 infants with community-acquired infection and 20.0% in 20 infants who acquired the infection during their birth hospitalization in the neonatal intensive care units (NICU) (nosocomial ARI) (p<0.001). In infants with community-acquired infection, Respiratory syncytial virus (RSV) (52.1%) and Rhinovirus/Enterovirus (RV/EV) (41.0%) were the most frequent detected pathogens. Coinfections were determined in one quarter of the infants, RSV-RV/EV being the most frequent combination. In infants with nosocomial infection, RV/EV, RSV or Parainfluenza-3 were detected as single pathogens. Most infants with community-acquired infection presented lower ARI (81.2%) while most infants in the NICU had upper ARI (55.0%). The median length of stay (LOS) in infants with community-acquired ARI was 4 days (IQR: 2-6). Positive infants with nosocomial infection had longer median LOS (71 days [IQR:42-99]) compared to negative infants (58 days [IQR: 49-71]) (p=0.507). Respiratory viruses were detected as the major causative agents of community-acquired infection in hospitalized infants <2-months old, RSV and RV/EV being the most frequently detected. Although a low pathogen positivity rate was observed in infants with nosocomial infection, they may prolong the LOS.
Assuntos
Infecções Respiratórias , Vírus , Criança , Hospitalização , Humanos , Lactente , Recém-Nascido , Vírus Sinciciais Respiratórios , Infecções Respiratórias/epidemiologia , Estudos RetrospectivosRESUMO
OBJECTIVE: Delineate risk factors associated with severe hypoxemia (O2 sat ≤87%) in infants and children younger than 2 years hospitalized with single pathogen HRV infection. STUDY DESIGN: Prospective study in a yearly catchment population of 56 560 children <2 years old between 2011 and 2013 in Argentina. All children with respiratory signs and O2 sat <93% on admission were included. HRV infections were identified by reverse transcriptase-polymerase chain reaction. Epidemiologic, clinical, viral, and immunological risk factors were assessed. RESULTS: Among 5012 hospitalized patients, HRV was detected as a single pathogen in 347 (6.92%) subjects. Thirty-two (9.2%) had life-threatening disease. Traditional risk factors for severe bronchiolitis did not affect severity of illness. HRV viral load, HRV groups, and type II and III interferons did not associate with severe hypoxemia. Interleukin-13 Levels in respiratory secretions at the time of admission (OR = 7.43 (3-18.4); P < 0.001 for IL-13 >10 pg/mL) predisposed to life-threatening disease. CONCLUSIONS: Targeted interventions against IL-13 should be evaluated to decrease severity of HRV illness in infancy and early childhood.
Assuntos
Bronquiolite/imunologia , Hipóxia/imunologia , Interleucina-13/imunologia , Infecções por Picornaviridae/imunologia , Infecções Respiratórias/imunologia , Rhinovirus , Argentina/epidemiologia , Bronquiolite/epidemiologia , Bronquiolite/virologia , Feminino , Hospitalização , Humanos , Hipóxia/epidemiologia , Hipóxia/virologia , Lactente , Recém-Nascido , Masculino , Infecções por Picornaviridae/epidemiologia , Infecções por Picornaviridae/virologia , Estudos Prospectivos , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologiaRESUMO
Rhinoviruses were detected as sole pathogens in 6 preterm infants who developed severe respiratory infections while hospitalized in a neonatal intensive care unit. We confirmed 2 nosocomial rhinovirus transmission episodes and describe the genetic diversity of rhinovirus strains that circulated simultaneously during a winter season.
Assuntos
Infecção Hospitalar/transmissão , Infecções por Picornaviridae/virologia , Rhinovirus/genética , Argentina , Infecção Hospitalar/virologia , Feminino , Variação Genética , Técnicas de Genotipagem/métodos , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Unidades de Terapia Intensiva Neonatal/estatística & dados numéricos , Masculino , Infecções por Picornaviridae/transmissão , Infecções Respiratórias/virologiaRESUMO
BACKGROUND: Although information about the incidence of viral respiratory illnesses and their associated cost can help health officials explore the value of interventions, data are limited from middle-income countries. METHODS: During 2008-2010, we conducted a prospective cohort study and followed ~1,800 Argentinian children aged ≤5 years to identify those children who were hospitalized or who sought care at an emergency room with any acute respiratory infection sign or symptom (e.g., rhinorrhea, cough, wheezing, tachypnea, retractions, or cyanosis). Respiratory samples were obtained for respiratory syncytial virus, influenza, parainfluenza, adenovirus, and metapneumovirus testing by immunofluorescence and for rhinovirus by real-time reverse transcription polymerase chain reaction. RESULTS: The incidence of respiratory syncytial virus (24/1000 children-years), human metapneumovirus (8/1000 children-years), and influenza (8/1000 children-years) illnesses was highest among hospitalized children aged <6 months and decreased among older children. In contrast, the incidence of rhinovirus was highest (12/1000 children-years) among those aged 6-23 months. In the emergency room, the incidence of rhinovirus was 459; respiratory syncytial virus 352; influenza 185; parainfluenza 177; metapneumovirus 130; and adenovirus 73/1,000 children-years. The total cost of hospitalization was a median of US$529 (Interquartile range, US$362-789). CONCLUSIONS: Our findings indicate that respiratory viruses, in particular rhinovirus, respiratory syncytial virus, metapneumovirus, and influenza may be associated with severe illness causing substantial economic burden.
Assuntos
Infecções Respiratórias/diagnóstico , Viroses/diagnóstico , Argentina/epidemiologia , Criança Hospitalizada , Pré-Escolar , Estudos de Coortes , Demografia , Serviço Hospitalar de Emergência/economia , Feminino , Humanos , Incidência , Lactente , Masculino , Metapneumovirus/genética , Metapneumovirus/isolamento & purificação , Microscopia de Fluorescência , Orthomyxoviridae/genética , Orthomyxoviridae/isolamento & purificação , Pacientes Ambulatoriais , Infecções por Paramyxoviridae/epidemiologia , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Vírus Sinciciais Respiratórios/genética , Vírus Sinciciais Respiratórios/isolamento & purificação , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Rhinovirus/genética , Rhinovirus/isolamento & purificação , Viroses/economia , Viroses/epidemiologiaRESUMO
Acute respiratory infections, which are commonly caused by viruses, are an important cause of morbidity and mortality in children. In Argentina, national surveillance programs for the detection of respiratory viruses are usually performed by using immunofluorescence (IF) assays, although it is well known that molecular methods are more sensitive. An automated multiplex PCR device, the FilmArray-Respiratory Panel (FilmArray-RP), can detect 17 viral and 3 bacterial pathogens in a closed system that requires only 5 min of hands-on time and 1h of instrumentation time. A total of 315 respiratory samples from children under 6 years of age suffering from acute respiratory infections, were studied by IF for 8 respiratory viruses and by RT-PCR for rhinoviruses. Later, these samples were tested by the FilmArray-RP. The positivity frequency obtained for the 9 viruses tested was 75% by IF/RT-PCR and 92% by the FilmArray-RP. The positive and negative percent agreement between both methods was 70.5% whereas the negative percent agreement was 99.6% (95% confidence interval:65.5-75.1 and 99.2-99.8 respectively). The FilmArray-RP allowed a higher positive diagnosis (97%) and detected other viruses such as coronavirus NL63, 229E, OC43, HKU1 (10%) and bocavirus (18%). In addition, this method identified multiple coinfections (39%) with 2, 3, 4 and up to 5 different viruses. At present, IF is still the most frequently used method in most Latin American countries for respiratory viruses diagnosis due to its low cost, its capability to process a high number of samples simultaneously and the fast determination of results for the most frequent viruses, which are available within 5h. However, the coming incorporation of molecular methods in routine procedures will significantly increase the diagnostic yield of these infections.
Assuntos
Reação em Cadeia da Polimerase Multiplex , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/virologia , Viroses/diagnóstico , Criança , Humanos , Estudos RetrospectivosRESUMO
Las infecciones respiratorias agudas producen una importante morbimortalidad y comúnmente son causadas por virus. En Argentina, los programas de vigilancia epidemiológica se basan en la detección de antígenos virales por inmunofluorescencia (IF), aunque es bien conocido que los métodos moleculares son más sensibles. El panel respiratorio (PR) FilmArray (PR-FilmArray) es un equipo comercial automatizado de PCR múltiples que detecta 17 virus respiratorios y 3 bacterias, en un sistema cerrado que requiere 5 min de procesamiento y una 1 h de instrumentación. Se evaluó un total de 315 muestras respiratorias de niños menores de 6 años con infecciones respiratorias agudas por IF para 8 virus respiratorios y por RT-PCR para rinovirus. Posteriormente, estas muestras se estudiaron con el PR-FilmArray. La frecuencia de positividad al considerar los 9 virus estudiados por IF y RT-PCR fue del 75 %; por PR-FilmArray fue del 92 %. El porcentaje de acuerdo positivo entre ambas metodologías fue del 70,5 % y el de acuerdo negativo fue del 99,6 % (intervalo de confianza 95 %: 65,5-75,1 y 99,2-99,8, respectivamente). El PR-FilmArray permitió obtener un mayor diagnóstico positivo (97 %) y detectó otros virus, como los coronavirus NL63, 229E, OC43 y HKU1 (10 %) y los bocavirus (18 %). Además, permitió identificar coinfecciones múltiples (39 %) con 2, 3, 4 y hasta 5 virus. Actualmente, la IF continúa siendo el método más utilizado en los países latinoamericanos para el diagnóstico de virus respiratorios por su bajo costo, por su capacidad para procesar un alto número de muestras simultáneamente y porque los resultados de los virus más frecuentes están disponibles en 5 h. Sin embargo, la futura incorporación de métodos moleculares aumentaría notablemente la capacidad diagnóstica.
Acute respiratory infections, which are commonly caused by viruses, are an important cause of morbidity and mortality in children. In Argentina, national surveillance programs for the detection of respiratory viruses are usually performed by using immunofluorescence (IF) assays, although it is well known that molecular methods are more sensitive. An automated multiplex PCR device, the FilmArray-Respiratory Panel (FilmArray-RP), can detect 17 viral and 3 bacterial pathogens in a closed system that requires only 5 min of hands-on time and 1 h of instrumentation time. A total of 315 respiratory samples from children under 6 years of age suffering from acute respiratory infections, were studied by IF for 8 respiratory viruses and by RT-PCR for rhinoviruses. Later, these samples were tested by the FilmArray-RP. The positivity frequency obtained for the 9 viruses tested was 75 % by IF/RT-PCR and 92 % by the FilmArray-RP. The positive and negative percent agreement between both methods was 70.5 % whereas the negative percent agreement was 99.6 % (95 % confidence interval:65.5-75.1 and 99.2-99.8 respectively). The FilmArray-RP allowed a higher positive diagnosis (97 %) and detected other viruses such as coronavirus NL63, 229E, OC43, HKU1 (10 %) and bocavirus (18 %). In addition, this method identified multiple coinfections (39 %) with 2, 3, 4 and up to 5 different viruses. At present, IF is still the most frequently used method in most Latin American countries for respiratory viruses diagnosis due to its low cost, its capability to process a high number of samples simultaneously and the fast determination of results for the most frequent viruses, which are available within 5 h. However, the coming incorporation of molecular methods in routine procedures will significantly increase the diagnostic yield of these infections.
Assuntos
Criança , Humanos , Reação em Cadeia da Polimerase Multiplex , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/virologia , Viroses/diagnóstico , Estudos RetrospectivosRESUMO
Las infecciones respiratorias agudas producen una importante morbimortalidad y comúnmente son causadas por virus. En Argentina, los programas de vigilancia epidemiológica se basan en la detección de antígenos virales por inmunofluorescencia (IF), aunque es bien conocido que los métodos moleculares son más sensibles. El panel respiratorio (PR) FilmArray (PR-FilmArray) es un equipo comercial automatizado de PCR múltiples que detecta 17 virus respiratorios y 3 bacterias, en un sistema cerrado que requiere 5 min de procesamiento y una 1 h de instrumentación. Se evaluó un total de 315 muestras respiratorias de niños menores de 6 años con infecciones respiratorias agudas por IF para 8 virus respiratorios y por RT-PCR para rinovirus. Posteriormente, estas muestras se estudiaron con el PR-FilmArray. La frecuencia de positividad al considerar los 9 virus estudiados por IF y RT-PCR fue del 75 %; por PR-FilmArray fue del 92 %. El porcentaje de acuerdo positivo entre ambas metodologías fue del 70,5 % y el de acuerdo negativo fue del 99,6 % (intervalo de confianza 95 %: 65,5-75,1 y 99,2-99,8, respectivamente). El PR-FilmArray permitió obtener un mayor diagnóstico positivo (97 %) y detectó otros virus, como los coronavirus NL63, 229E, OC43 y HKU1 (10 %) y los bocavirus (18 %). Además, permitió identificar coinfecciones múltiples (39 %) con 2, 3, 4 y hasta 5 virus. Actualmente, la IF continúa siendo el método más utilizado en los países latinoamericanos para el diagnóstico de virus respiratorios por su bajo costo, por su capacidad para procesar un alto número de muestras simultáneamente y porque los resultados de los virus más frecuentes están disponibles en 5 h. Sin embargo, la futura incorporación de métodos moleculares aumentaría notablemente la capacidad diagnóstica.(AU)
Acute respiratory infections, which are commonly caused by viruses, are an important cause of morbidity and mortality in children. In Argentina, national surveillance programs for the detection of respiratory viruses are usually performed by using immunofluorescence (IF) assays, although it is well known that molecular methods are more sensitive. An automated multiplex PCR device, the FilmArray-Respiratory Panel (FilmArray-RP), can detect 17 viral and 3 bacterial pathogens in a closed system that requires only 5 min of hands-on time and 1 h of instrumentation time. A total of 315 respiratory samples from children under 6 years of age suffering from acute respiratory infections, were studied by IF for 8 respiratory viruses and by RT-PCR for rhinoviruses. Later, these samples were tested by the FilmArray-RP. The positivity frequency obtained for the 9 viruses tested was 75 % by IF/RT-PCR and 92 % by the FilmArray-RP. The positive and negative percent agreement between both methods was 70.5 % whereas the negative percent agreement was 99.6 % (95 % confidence interval:65.5-75.1 and 99.2-99.8 respectively). The FilmArray-RP allowed a higher positive diagnosis (97 %) and detected other viruses such as coronavirus NL63, 229E, OC43, HKU1 (10 %) and bocavirus (18 %). In addition, this method identified multiple coinfections (39 %) with 2, 3, 4 and up to 5 different viruses. At present, IF is still the most frequently used method in most Latin American countries for respiratory viruses diagnosis due to its low cost, its capability to process a high number of samples simultaneously and the fast determination of results for the most frequent viruses, which are available within 5 h. However, the coming incorporation of molecular methods in routine procedures will significantly increase the diagnostic yield of these infections.(AU)
RESUMO
Acute respiratory infections, which are commonly caused by viruses, are an important cause of morbidity and mortality in children. In Argentina, national surveillance programs for the detection of respiratory viruses are usually performed by using immunofluorescence (IF) assays, although it is well known that molecular methods are more sensitive. An automated multiplex PCR device, the FilmArray-Respiratory Panel (FilmArray-RP), can detect 17 viral and 3 bacterial pathogens in a closed system that requires only 5min of hands-on time and 1h of instrumentation time. A total of 315 respiratory samples from children under 6 years of age suffering from acute respiratory infections, were studied by IF for 8 respiratory viruses and by RT-PCR for rhinoviruses. Later, these samples were tested by the FilmArray-RP. The positivity frequency obtained for the 9 viruses tested was 75
by IF/RT-PCR and 92
by the FilmArray-RP. The positive and negative percent agreement between both methods was 70.5
whereas the negative percent agreement was 99.6
(95
confidence interval:65.5-75.1 and 99.2-99.8 respectively). The FilmArray-RP allowed a higher positive diagnosis (97
) and detected other viruses such as coronavirus NL63, 229E, OC43, HKU1 (10
) and bocavirus (18
). In addition, this method identified multiple coinfections (39
) with 2, 3, 4 and up to 5 different viruses. At present, IF is still the most frequently used method in most Latin American countries for respiratory viruses diagnosis due to its low cost, its capability to process a high number of samples simultaneously and the fast determination of results for the most frequent viruses, which are available within 5h. However, the coming incorporation of molecular methods in routine procedures will significantly increase the diagnostic yield of these infections.
RESUMO
BACKGROUND: Human rhinoviruses (HRV) are recognized as a cause of upper and lower acute respiratory infections (ARI). The circulating species and their clinical impact were not described in Argentina. OBJECTIVES: To describe the molecular epidemiology of HRV in children and to determine the association of HRV species with outcome and severity. STUDY DESIGN: Hospitalized and outpatients children <6 years old with ARI without comorbidities (n=620) were enrolled (2008-2010). Demographic, clinical data and outcome were analyzed. HRV were identified by RT-PCR. Phylogenetic analysis and demographic reconstruction for HRV were performed in selected samples. RESULTS: HRV were detected in 252/620 (40.6%) of children; 8.5% in viral coinfection. Bronchiolitis (55%) and pneumonia (13%) were the most frequent clinical diagnosis. Of 202 inpatients with HRV: 72% required oxygen supplementation, 11% intensive care unit and 3% mechanical ventilation. HRV were identified as a risk factor for hospitalization (OR: 2.47). All three HRV species were detected being HRV-A (55%) and HRV-C (43%) the most frequent; HRV-B was infrequent (2%). Of 44 sequenced HRV, 30 genotypes were detected. Seven of them were the most prevalent and circulated during limited periods of time. The demographic reconstruction revealed a constant population size and a high turnover rate of genotypes. Demographic and clinical outcome were similar for HRV-A and HRV-C infections. CONCLUSION: This study highlights the clinical impact of HRV in children without comorbidities as a cause of lower ARI and hospitalization. The high frequency of HRV infections may be associated with the simultaneous circulation of genotypes and their high turnover rate.
Assuntos
Variação Genética , Infecções por Picornaviridae/patologia , Infecções por Picornaviridae/virologia , Infecções Respiratórias/patologia , Infecções Respiratórias/virologia , Rhinovirus/classificação , Rhinovirus/genética , Argentina , Pré-Escolar , Análise por Conglomerados , Estudos Transversais , Feminino , Genótipo , Humanos , Lactente , Pacientes Internados , Masculino , Epidemiologia Molecular , Dados de Sequência Molecular , Pacientes Ambulatoriais , Filogenia , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rhinovirus/isolamento & purificação , Análise de Sequência de DNA , Homologia de Sequência , Resultado do TratamentoRESUMO
BACKGROUND: Between September 2000 and November 2005, approximately 10% of the retrospectively examined human adenovirus (HAdV)-positive pediatric cases of acute respiratory disease (ARD) requiring hospitalization at the Hospital Nacional de Pediatria Juan P. Garrahan in Buenos Aires, Argentina, were found to have a HAdV-B2 infection. OBJECTIVE: To characterize genetically and antigenically the HAdV-B2 virus isolates. STUDY DESIGN: Restriction enzyme analysis (REA), hexon and fiber gene sequencing and virus neutralization assays (VN) were carried out on 8 HAdV-B2 respiratory virus isolates. RESULTS: REA showed that the 8 examined HAdV-B2 virus isolates were HAdV11, belonging to two genomic variants: HAdV11a and a BclI variant of HAdV11c which we designated 11c4. Molecular analysis of the hexon genes showed that both REA variants had a HAdV11-like hexon gene. Confirming previous reports, the 7 HAdV11a virus isolates were found to have HAdV14-like fiber genes and therefore are HAdV H11/F14. The fiber gene of the HAdV11c4 virus isolates most closely resembled that of various strains of HAdV7. In VN assays, the 4 tested HAdV11a strains were serotyped as HAdV11-14. The HAdV11c4 strain was serotyped as HAdV11 but also showed a weak but significant reactivity with antiserum to HAdV7. Compared with the other HAdV-positive cases in our study, infection with HAdV11 caused a similarly severe disease. CONCLUSIONS: Our results provide evidence to the long term world-wide circulation of HAdV H11/F14 as a causative agent of ARD. Combined, our molecular and serology data support the rationale to base the molecular typing and designation of recombinant viruses on the sequences of the hexon and fiber genes.