Comparison of the direct fluorescence assay and real-time polymerase chain reaction for the detection of influenza virus A and B in immunocompromised patients.

OBJECTIVE: This study evaluated the diagnostic performance of two methods for the detection of influenza virus in immunocompromised transplant patients. METHODS: A total of 475 respiratory samples, 236 from patients in a hematopoietic stem cell transplantation program and 239 from kidney transplant patients, were analyzed by a direct fluorescence assay and the Centers for Disease Control real-time polymerase chain reaction protocol for influenza A and B detection. RESULTS: Influenza detection using either method was 7.6% in the hematopoietic stem cell transplant group and 30.5% in the kidney transplant patient group. Influenza detection by real-time polymerase chain reaction yielded a higher positive rate compared with fluorescence than that reported by other studies, and this difference was more pronounced for influenza A. The fluorescence assay sensitivity, specificity, positive and negative predictive values, and kappa coefficient were 17.6%, 100%, 1, 0.83, and 0.256, respectively, and lower detection rates occurred in the kidney transplant patients. CONCLUSIONS: The real-time polymerase chain reaction performance and the associated turnaround time for a large number of samples support the choice of this method for use in different routine diagnostic settings and influenza surveillance in high-risk patients.

Comparison of the direct fluorescence assay and real-time polymerase chain reaction for the detection of influenza virus A and B in immunocompromised patients

OBJECTIVE: This study evaluated the diagnostic performance of two methods for the detection of influenza virus in immunocompromised transplant patients. METHODS: A total of 475 respiratory samples, 236 from patients in a hematopoietic stem cell transplantation program and 239 from kidney transplant patients, were analyzed by a direct fluorescence assay and the Centers for Disease Control real-time polymerase chain reaction protocol for influenza A and B detection. RESULTS: Influenza detection using either method was 7.6% in the hematopoietic stem cell transplant group and 30.5% in the kidney transplant patient group. Influenza detection by real-time polymerase chain reaction yielded a higher positive rate compared with fluorescence than that reported by other studies, and this difference was more pronounced for influenza A. The fluorescence assay sensitivity, specificity, positive and negative predictive values, and kappa coefficient were 17.6%, 100%, 1, 0.83, and 0.256, respectively, and lower detection rates occurred in the kidney transplant patients. CONCLUSIONS: The real-time polymerase chain reaction performance and the associated turnaround time for a large number of samples support the choice of this method for use in different routine diagnostic settings and influenza surveillance in high-risk patients. .

Human adenovirus detection among immunocompetent and immunocompromised patients presenting acute respiratory infection.

INTRODUCTION: Human adenoviruses (HAdV) play an important role in the aetiology of severe acute lower respiratory infection, especially in immunocompromised individuals. The aim of the present study was to detect HAdV using two different methods, direct fluorescence assay (DFA) and nested polymerase chain reaction (nested PCR), in samples collected from patients with acute groups from 2001 to 2010: 139 adult emergency room patients (ERP); 205 health care workers (HCW); 69 renal transplant outpatients(RTO); and 230 patients in a haematopoietic stem cell transplantation program (HSCT). RESULTS: Adenovirus was detected in 13.2% of the 643 patients tested by DFA and/or PCR: 6/139 (4.3%) adults in the ERP group, 7/205 (3.4%) in the HCW group, 4/69 (5.8%) in the RTO group and 68/230 (29.5%) in the HSCT patient group. Nested PCR had a higher detection rate (10%) compared with the DFA test (3.8%) (p<0.001). HSCT patients exhibited a significantly higher rate of HAdV infection. CONCLUSIONS: The adenovirus detection rate of the nested PCR assay was higher than that of the DFA test. However, the use of molecular methods in routine diagnostic laboratory work should be evaluated based on the specific circumstances of individual health services.

Human adenovirus detection among immunocompetent and immunocompromised patients presenting acute respiratory infection

INTRODUCTION: Human adenoviruses (HAdV) play an important role in the etiology of severe acute lower respiratory infection, especially in immunocompromised individuals. The aim of the present study was detect the HAdV through different methods: direct fluorescence assay (DFA) and nested-polymerase chain reaction (PCR-nested) from patients with acute respiratory infection (ARI) up to 7 days of symptoms onset. METHODS: Samples (n=643) were collected from different risk groups during from 2001 to 2010: 139 adults attended in an Emergency Room Patients (ERP); 205 health care workers (HCW); 69 from Renal Transplant Outpatients (RTO); 230 patients in hematopoietic stem cell transplantation (HSCT) program. RESULTS: Among all patients (n=643) adenovirus was detected on 13.2% by DFA and/or PCR: 6/139 (4.3%) adults from ERP, 7/205 (3.4%) from HCW samples, 4/69 (5.8%) from RTO and 68/230 (29.5%) from HSCT patients. Nested PCR showed higher detection (10%) compared to DFA test (3.8%) (p < 0.001). HSCT patients presented significantly higher prevalence of HAdV infection. CONCLUSIONS: Adenovirus detection through nested-PCR assay was higher. However the inclusion of molecular method in laboratorial routine diagnostic should be evaluated considering the reality of each specific health service. .

A survey strategy for human respiratory syncytial virus detection among haematopoietic stem cell transplant patients: epidemiological and methodological analysis.

Human respiratory syncytial virus (HRSV) causes severe infections among children and immunocompromised patients. We compared HRSV infections among Haematopoietic Stem Cell Transplant program (HSCT) patients and children using direct immunofluorescence (DFA), point-of-care RSV Bio Easy® and a polymerase chain reaction (PCR) assay. Overall, 102 samples from HSCT patients and 128 from children obtained positivity rate of 18.6% and 14.1% respectively. PCR sensitivity was highest mainly on samples collected after five days of symptoms onset. A combination of both DFA and reverse transcriptase-PCR methods for HSCT high-risk patients is the best diagnostic flow for HRSV diagnosis among these patients.

A survey strategy for human respiratory syncytial virus detection among haematopoietic stem cell transplant patients: epidemiological and methodological analysis

Human respiratory syncytial virus (HRSV) causes severe infections among children and immunocompromised patients. We compared HRSV infections among Haematopoietic Stem Cell Transplant program (HSCT) patients and children using direct immunofluorescence (DFA), point-of-care RSV Bio Easy® and a polymerase chain reaction (PCR) assay. Overall, 102 samples from HSCT patients and 128 from children obtained positivity rate of 18.6% and 14.1% respectively. PCR sensitivity was highest mainly on samples collected after five days of symptoms onset. A combination of both DFA and reverse transcriptase-PCR methods for HSCT high-risk patients is the best diagnostic flow for HRSV diagnosis among these patients.

Surveillance of influenza A H1N1 2009 among school children during 2009 and 2010 in São Paulo, Brazil.

INTRODUCTION: Influenza A H1N1 2009 is associated with a high morbidity rate among children around the world, including Brazil. This survey was conducted on samples of symptomatic children (< 12 years) to investigate the influenza virus as the etiological agent of respiratory infections in a day care school in a health facility during the first and second pandemic wave of H1N1 (2009-2010) in São Paulo, Brazil. METHODS: Influenza infections were determined by real-time PCR in 34% (47/137) of children with a median age of 5 years (8 months - 12 years), from June to October 2009 and in 16% (14/85) of those with median age of 6 years (1-12 years), from March to November 2010. RESULTS: In general, most positive cases (64%) occurred in children aged 5-12 years, this age group was significantly the most affected (39.8%, p = 0.001, OR = 8.3, CI 95% 1.9-36.9). Wheezing was reported by 31% (19/61) and dyspnea by 23% (14/61) of the studied patients. An outbreak of influenza H1N1 with an attack rate of 35.7% among children (median age 6 years) was documented in April 2010, before the vaccination campaign against the pandemic virus was extended for children up to 5 years in Brazil. CONCLUSIONS: Therefore, the study reinforces the recommendation to immunize school children to reduce the incidence of the disease.

Surveillance of influenza A H1N1 2009 among school children during 2009 and 2010 in São Paulo, Brazil / Vigilância de influenza A H1N1 2009 entre crianças em idade escolar durante os anos de 2009 e 2010 em São Paulo, Brasil

INTRODUCTION: Influenza A H1N1 2009 is associated with a high morbidity rate among children around the world, including Brazil. This survey was conducted on samples of symptomatic children (< 12 years) to investigate the influenza virus as the etiological agent of respiratory infections in a day care school in a health facility during the first and second pandemic wave of H1N1 (2009-2010) in São Paulo, Brazil. METHODS: Influenza infections were determined by real-time PCR in 34% (47/137) of children with a median age of 5 years (8 months - 12 years), from June to October 2009 and in 16% (14/85) of those with median age of 6 years (1-12 years), from March to November 2010. RESULTS: In general, most positive cases (64%) occurred in children aged 5-12 years, this age group was significantly the most affected (39.8%, p = 0.001, OR = 8.3, CI 95% 1.9-36.9). Wheezing was reported by 31% (19/61) and dyspnea by 23% (14/61) of the studied patients. An outbreak of influenza H1N1 with an attack rate of 35.7% among children (median age 6 years) was documented in April 2010, before the vaccination campaign against the pandemic virus was extended for children up to 5 years in Brazil. CONCLUSIONS: Therefore, the study reinforces the recommendation to immunize school children to reduce the incidence of the disease.

INTRODUÇÃO: Influenza A H1N1 2009 está associado com uma alta taxa de morbidade entre crianças ao redor do mundo, incluindo o Brasil. Esta pesquisa foi realizada em amostras de crianças sintomáticas (< 12 anos) em uma creche escola para filhos de funcionários do hospital durante a primeira e segunda onda pandêmica (2009-2010) em São Paulo, Brasil. MÉTODOS: Infecções pelo vírus influenza foram determinadas por PCR em tempo real em 34% (47/137) em crianças com idade mediana de 5 anos (8 meses -12 anos), entre junho e outubro de 2009 e em 16% (14/85) daquelas com mediana de idade de 6 anos (1-12 anos), de março a novembro de 2010. RESULTADOS: Em geral, a maioria dos casos positivos (64%) ocorreu em crianças com idade entre 5-12 anos, esta faixa etária foi significativamente a mais afetada (39,8%, p = 0,001, OR = 8,3, CI 95%: 1,9-36,9). Chiado foi relatado em 31% (19/61) e dispnéia em 23% (14/61) dos pacientes estudados. Um surto de gripe H1N1 com uma taxa de ataque de 35,7% entre as crianças (mediana de idade de 6 anos) foi documentado em abril de 2010, antes da extensão da campanha de vacinação contra o vírus pandêmico para crianças até 5 anos no Brasil. CONCLUSÕES: Portanto, neste estudo reforça a recomendação para imunizar crianças em idade escolar para reduzir a incidência da doença.

Respiratory viral coinfection among hospitalized patients with H1N1 2009 during the first pandemic wave in Brazil.

Influenza A coinfections with other respiratory viruses were investigated in 25.8% (41/159) of the samples from patients hospitalized in 2009 at our University Hospital. Out of the 41 influenza A cases, nine cases (21.9%) were coinfected with other viruses, with a similar frequency among children and adults (p = 0.47), and seasonal influenza cases were more prevalent than H1N1 2009 influenza virus. Adenovirus was the most frequently detected (4/9) among coinfected cases. Coinfection was not associated with higher morbidity or mortality (p = 0.75).

Occurrence of influenza among patients hospitalized for suspicion of influenza A (H1N1) infection in 2010 at a sentinel hospital in São Paulo, Brazil.

In 2010, 96 patients suspected of being infected with the influenza A (H1N1) virus were hospitalized at the Hospital São Paulo, located in the city of São Paulo, Brazil. Of those 96 patients, 4 (4.2%) were found to be infected with influenza A virus-3 with influenza A (H1N1) and 1 with seasonal influenza A-and 2 patients (2.1%) were found to be infected with influenza B virus. Most (63.5%) of the suspected cases occurred in children, as did half of the positive cases. The second wave of influenza A (H1N1) infection was weaker in São Paulo. The decrease in the number of hospitalizations for H1N1 infection in 2010 might be attributable to vaccination.

Casos confirmados de influenza em pacientes hospitalizados com suspeita de infecção por influenza A (H1N1) em 2010 em um hospital sentinela na cidade de São Paulo / Occurrence of influenza among patients hospitalized for suspicion of influenza A (H1N1) infection in 2010 at a sentinel hospital in São Paulo, Brazil

Em 2010, 96 pacientes com suspeita de infecção por influenza A (H1N1) foram hospitalizados no Hospital São Paulo, na cidade de São Paulo (SP). Desses, 4 pacientes (4,2 por cento) foram diagnosticados com influenza A - 3 com influenza A (H1N1) e 1 com influenza sazonal - e 2 pacientes (2,1 por cento) foram diagnosticados com influenza B. A maioria dos casos suspeitos (63,5 por cento) e metade dos casos positivos ocorreram em crianças. A segunda onda de influenza A (H1N1) foi mais fraca em São Paulo. A vacinação pode ter contribuído para a redução das internações devido a essa infecção em 2010.

In 2010, 96 patients suspected of being infected with the influenza A (H1N1) virus were hospitalized at the Hospital São Paulo, located in the city of São Paulo, Brazil. Of those 96 patients, 4 (4.2 percent) were found to be infected with influenza A virus-3 with influenza A (H1N1) and 1 with seasonal influenza A-and 2 patients (2.1 percent) were found to be infected with influenza B virus. Most (63.5 percent) of the suspected cases occurred in children, as did half of the positive cases. The second wave of influenza A (H1N1) infection was weaker in São Paulo. The decrease in the number of hospitalizations for H1N1 infection in 2010 might be attributable to vaccination.

Respiratory virus infections among hospitalized patients with suspected influenza A H1N1 2009 virus during the first pandemic wave in Brazil.

INTRODUCTION: During the first pandemic wave of the influenza A H1N1 2009 virus, morbidity was particularly high in Brazil. Hospitalizations resulting from severe respiratory disease due to suspected influenza-like illness created an opportunity to identify other respiratory viruses causing lower respiratory infections. OBJECTIVE: The purpose of this study was to assess viral etiologies among samples collected during the first pandemic wave of H1N1 2009 from hospitalized patients with suspected cases in a Brazilian Sentinel Hospital. PATIENTS AND METHODS: Viral etiologies were investigated in samples from 98 children and 61 adults with fever, cough and dyspnea who were admitted to São Paulo Sentinel Hospital with suspected H1N1 infection. RESULTS: From August to November 2009, in 19.5% (31/159) of the samples 2009 H1N1 virus was detected with 23% (14/61) in adults (median age 25 years, range: 14-55 years) and 18.4% (17/92) in children (median age 5 years, range: 4 months - 11 years). Among the negative samples, a wide range of causative etiologic agents was identified. Human rhinovirus was the most frequent virus (23.91%) in children and human metapneumovirus (11.48%) was the second most frequent in adults, following 2009 H1N1 virus (22.95%). CONCLUSION: These data highlight the need to diagnose other viral infections that can co-circulate with influenza and may have been neglected by physicians as causes of severe respiratory diseases.

Respiratory virus infections among hospitalized patients with suspected influenza A H1N1 2009 virus during the first pandemic wave in Brazil

INTRODUCTION: During the first pandemic wave of the influenza A H1N1 2009 virus, morbidity was particularly high in Brazil. Hospitalizations resulting from severe respiratory disease due to suspected influenza-like illness created an opportunity to identify other respiratory viruses causing lower respiratory infections. OBJECTIVE: The purpose of this study was to assess viral etiologies among samples collected during the first pandemic wave of H1N1 2009 from hospitalized patients with suspected cases in a Brazilian Sentinel Hospital. PATIENTS AND METHODS: Viral etiologies were investigated in samples from 98 children and 61 adults with fever, cough and dyspnea who were admitted to São Paulo Sentinel Hospital with suspected H1N1 infection. RESULTS: From August to November 2009, in 19.5 percent (31/159) of the samples 2009 H1N1 virus was detected with 23 percent (14/61) in adults (median age 25 years, range: 14-55 years) and 18.4 percent (17/92) in children (median age 5 years, range: 4 months - 11 years). Among the negative samples, a wide range of causative etiologic agents was identified. Human rhinovirus was the most frequent virus (23.91 percent) in children and human metapneumovirus (11.48 percent) was the second most frequent in adults, following 2009 H1N1 virus (22.95 percent). CONCLUSION: These data highlight the need to diagnose other viral infections that can co-circulate with influenza and may have been neglected by physicians as causes of severe respiratory diseases.

Increased exhalation of hydrogen peroxide in healthy subjects following cigarette consumption

CONTEXT: Increased hydrogen peroxide has been described in the expired breath condensate (H2O2-E) of several lung conditions, such as acute respiratory distress syndrome, chronic obstructive pulmonary disease and asthma. This technique has been advocated as being a simple method for documenting airway inflammation. OBJECTIVE: To evaluate H2O2-E in healthy cigarette smokers, and to determine the acute effects of the consumption of one cigarette on H2O2-E levels. TYPE OF STUDY: Prospective, controlled trial. SETTING: A pulmonary function laboratory in a University Hospital. PARTICIPANTS: Two groups of healthy volunteers: individuals who had never smoked (NS; n=10; 4 men; age = 30.6 Ý 6.2 years) and current cigarette smokers (S; n=12; 7 men; age = 38.7 Ý 9.8). None of the volunteers had respiratory symptoms and all showed normal spirometric tests. INTERVENTION: Expired air was collected from all volunteers through a face mask and a plastic collecting system leading into a flask with dry ice and pure ethanol. Samples from the group S were collected twice, before and half an hour after the combustion of one cigarette. MAIN MEASUREMENTS: Expired hydrogen peroxide using the Gallati and Pracht method. RESULTS: The S and NS groups showed comparable levels of H2O2-E at basal conditions [NS = 0.74 muM (DP 0.24) vs. S = 0.75 muM (DP 0.31)]. The smokers showed a significant increase in H2O2-E levels half an hour after the consumption of only one cigarette [0.75 muM (DP 0.31) vs. 0.95 muM (DP 0.22)]. CONCLUSION: The present results are consistent with the concept that smokers increase oxidative stress with elevated production of reactive oxygen species, contributing to the development of smoking-related disorders