Débora Natalia Marcone

Viral etiology of acute respiratory infections in hospitalized and outpatient children in Buenos Aires, Argentina.

Objectives: To determine and compare the viral frequency, seasonality and clinical-demographic features in 2 groups of children (hospitalized versus outpatients) with acute respiratory infections. Material and Methods: A cross-sectional, descriptive study was performed from 2008 to 2010 in 620 children <6 years of age with acute respiratory infection. Respiratory samples were studied for classical respiratory viruses by immunofluorescence and for human rhinoviruses (HRV) by real-time reverse transcription polymerase chain reaction. Clinical and demographic data were recorded. Results: Viral detection by immunofluorescence was 48% in 434 inpatients and 37% in 186 outpatients. Viral diagnosis increased to 83% and 62%, respectively, when testing for HRV. HRV (41%) and respiratory syncytial virus (RSV) (27%) were most common viruses identified, followed by metapneumovirus (9%), influenza A and parainfluenza (3%), adenovirus and influenza B (2%). HRV frequency was significantly higher in hospitalized patients (47%) than in outpatients (27%) (P < 0.001). Coinfection was detected in 12% of hospitalized and 4% of outpatients (P < 0.031). HRV and adenovirus circulated throughout the entire year. RSV, influenza A and B predominated in winter, whereas metapneumovirus and parainfluenza predominated in spring. Of 362 patients with bronchiolitis, 84% had a virus identified; HRV (42%) and RSV (38%) were predominant. Of 77 patients with pneumonia, 84% had a virus detected with HRV (43%) and RSV (29%) predominating. Conclusions: HRV were significant pathogens associated with bronchiolitis and pneumonia, especially in hospitalized patients. Both, HRV and coinfections, were risk factors for hospitalization. These findings support the importance of including HRV detection in children with acute respiratory infection.

Pandemic (H1N1) 2009 cases, Buenos Aires, Argentina.

To determine clinical and virologic characteristics of pandemic (H1N1) 2009 in Buenos Aires, Argentina, we conducted real-time reverse transcription–PCR on samples from patients with influenza-like illness, June 11–30, 2009. Of 513 patients tested, 54% were positive for influenza virus subtype H1N1. Infection rate was lowest for patients ≥60 years of age.

[Acute viral respiratory infection in children under 5 years: Epidemiological study in two centers in Buenos Aires, Argentina].

INTRODUCTION Acute respiratory infections (ARI) are a major cause of hospitalization particularly at the extreme ages of life. OBJECTIVES To determine the viral diagnosis in children < 5 years old with ARI, seasonality, clinical and epidemiological characteristics. POPULATION AND METHODS A cross-sectional, descriptive and multicenter study was performed. Respiratory samples from patients with ARI hospitalized or outpatients with < 5 days of respiratory symptoms from June 2008 to May 2009 were studied for respiratory viruses. Clinical and demographic data were recorded. RESULTS A total of 347 patients were enrolled: 234 hospitalized and 112 outpatients. Hospitalized patients were younger compared with outpatients (8 vs. 19 months, p <0.001) and presented more frequently bronchiolitis (74% hospitalized vs. 24% outpatients) and pneumonia (14% hospitalized vs. 5% outpatients). Age was statistically associated with hospitalization (p= 0.01). Viral diagnosis was achieved in 81% hospitalized and 57% of outpatients. Rhinovirus was the most frequent followed by respiratory syncytial virus. The rest of respiratory viruses were observed with lower frequency. Viral circulation was observed throughout the whole year. The median length of stay was 3.5 days. Intensive care was required in 11% of hospitalized patients and 3% required mechanical ventilation. No deaths were recorded. CONCLUSIONS The use of viral diagnostic techniques allowed the identification of an etiologic agent in most of the hospitalized patients and more than half of outpatients. The addition of RT-PCR for rhinovirus, allowed the identification of this etiologic agent. Viral circulation was observed throughout the whole year. Hospitalized patients presented bronchiolitis and pneumonia more frequently than outpatients.

Diagnóstico de virus respiratorios utilizando un sistema automatizado de PCR múltiples (FilmArray) y su comparación con métodos convencionales

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. Resumen 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 5min de procesamiento y una 1h 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 5h. Sin embargo, la futura incorporación de métodos moleculares aumentaría notablemente la capacidad diagnóstica. Abstract 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.

Genetic diversity and clinical impact of human rhinoviruses in hospitalized and outpatient children with acute respiratory infection, Argentina

Abstract 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.

Interleukin-13 associates with life-threatening rhinovirus infections in infants and young children

Delineate risk factors associated with severe hypoxemia (O2 sat ≤87%) in infants and children younger than 2 years hospitalized with single pathogen HRV infection.

Clinical impact of rapid molecular detection of respiratory pathogens in patients with acute respiratory infection

Abstract Background Acute respiratory infections (ARI) are a leading cause of morbidity and mortality worldwide. There is a need to demonstrate the clinical impact of using the new, rapid and sensitive molecular assays in prospectively designed studies. Objectives To study the impact on medical management of a rapid molecular assay in patients with respiratory infections. Study design A prospective, randomized, non-blinded study was performed in patients presenting to the Emergency Department during two respiratory seasons (2016–2017). Diagnosis was performed by FilmArray Respiratory Panel (FilmArray-RP) or by immunofluorescence assay (IFA). Results A total of 432 patients (156 children and 276 adults) were analyzed. Diagnosis with FilmArray-RP was associated with significant changes in medical management including withholding antibiotic prescriptions (OR:15.52, 95%CI:1.99–120.83 in adults and OR:12.23, 95%CI:1.56–96.09 in children), and reduction in complementary studies in children (OR:9.64, 95%CI:2.13–43.63) compared to IFA. Decrease in oseltamivir prescriptions was significantly higher in adults in the FilmArray-RP group (p = 0.042; OR:1.19, 95%CI:0.51-2.79) compared to adults managed with IFA. Diagnostic yield was significantly higher by FilmArray-RP (81%) than by IFA (31%)(p < 0.001). The median time from sample collection to reporting was 1 h 52 min by FilmArray-RP and 26 h by IFA (p < 0.001). Conclusions The high respiratory viruses’ detection rate and availability of results within two hours when using FilmArray-RP were associated with decreases in antibiotic prescriptions and complementary studies and more accurate use of oseltamivir.