ABSTRACT
Considering the great lethality and sequels caused by meningitis, rapid diagnosis and prompt treatment initiation have a great impact on patient outcome. Here, we developed a multiplex-PCR for simultaneous detection of the four most prevalent bacterial pathogens directly in CSF samples. The multiplex-PCR was designed to detect the following genes: fbsA (Streptococcus agalactiae), lytA (Streptococcus pneumoniae), crtA (Neisseria meningitidis), p6 (Haemophilus influenzae), and 16S rRNA (any bacterial agent). The multiplex-PCR showed a DNA detection limit of 1 pg/µL. Among 447 CSF samples tested, 40 were multiplex-PCR positive, in which 27 and 13 had positive and negative bacterial culture, respectively. Our multiplex-PCR is fast, reliable, and easily implementable into a laboratory routine for bacterial meningitis confirmation, especially for patients who previously started antimicrobial therapy. Our molecular approach can substantially improve clinical diagnosis and epidemiological measures of meningitis disease burden.
Subject(s)
Cerebrospinal Fluid/microbiology , Haemophilus influenzae/genetics , Meningitis, Bacterial/diagnosis , Neisseria meningitidis/genetics , Streptococcus agalactiae/genetics , Streptococcus pneumoniae/genetics , Adolescent , Adult , Aged , Child , Child, Preschool , Female , Haemophilus influenzae/isolation & purification , Humans , Infant , Infant, Newborn , Male , Meningitis, Bacterial/microbiology , Multiplex Polymerase Chain Reaction/methods , Neisseria meningitidis/isolation & purification , Streptococcus agalactiae/isolation & purification , Streptococcus pneumoniae/isolation & purificationABSTRACT
Human respiratory syncytial virus is the main cause of respiratory infections in infants. Several HRSV genotypes have been described. Goals. To describe the main genotypes that caused infections in São Paulo (2013-2015) and to analyze their clinical/epidemiological features. Methods. 94 infants (0-6 months) with bronchiolitis were studied. Clinical/epidemiological information was collected; a search for 16 viruses in nasopharyngeal secretion (PCR-real-time and conventional, sequencing, and phylogenetic analyses) was performed. Results. The mean age was 2.4 m; 48% were male. The mean length of hospital stay was 4.4 d (14% in the Intensive Care Unit). The positive rate of respiratory virus was 98.9%; 73 cases (77.6%) were HRSV (76,7% HRSVA). HRSVA formed three clusters: ON1 (n = 34), NA1 (n = 1), and NA2 (n = 4). All HRSVB were found to cluster in the BA genotype (BA9-n = 10; BA10-n = 3). Clinical analyses showed no significant differences between the genotype AON1 and other genotypes. Conclusion. This study showed a high rate of HRSV detection in bronchiolitis. HRSVA ON1, which has recently been described in other countries and has not been identified in previous studies in the southeast region of Brazil, was predominant. The clinical characteristics of the infants that were infected with AON1 were similar to infants with infections by other genotypes.
Subject(s)
Bronchiolitis, Viral/genetics , Genotype , Hospitalization , Respiratory Syncytial Virus Infections/genetics , Respiratory Syncytial Virus, Human/genetics , Brazil , Bronchiolitis, Viral/therapy , Female , Humans , Infant , Infant, Newborn , Male , Real-Time Polymerase Chain Reaction , Respiratory Syncytial Virus Infections/therapy , Respiratory Syncytial Virus, Human/isolation & purificationABSTRACT
Abstract Objective: The aim of this study was to evaluate the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA) as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. Methods: The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. Results: From 313 positive samples by immunofluorescence assays, 282 (90%) were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue® RSV Test and viral load or specific strain. The QuickVue® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. Conclusions: This study demonstrated that the QuickVue® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics.
Resumo Objetivo: Avaliar o teste QuickVue® RSV Test Kit (QUIDEL Corp, CA, EUA) para o diagnóstico rápido do vírus sincicial respiratório em crianças com doença respiratória aguda, comparandoo com a imunofluorescência indireta como padrão ouro. Visto que, no Brasil, testes rápidos para detecção de antígenos para vírus sincicial respiratório não são rotineiramente utilizados como ferramenta de diagnóstico, exceto para Dengue e Influenza. Métodos: Um total de 486 amostras de aspirado de nasofaringe de crianças menores de 5 anos com doença respiratória aguda, coletadas entre dezembro de 2013 e agosto de 2014, foram analisadas por imunofluorescência e pelo teste QuickVue®. Amostras com resultados discordantes entre os métodos foram submetidas a PCR em tempo real e sequenciamento. Resultados: Das 313 amostras positivas por IFI, 282 foram positivas no teste rápido (90%), 2 amostras foram positivas apenas no teste rápido (0.6%), 33 apenas na imunofluorescência (10.5%) e 171 foram negativas em ambos os métodos. As 35 amostras com resultados discordantes foram testadas por PCR em tempo real, sendo que duas que foram positivas apenas no teste rápido e 5 apenas na imunofluorescência confirmaram-se positivas. Não houve relação entre a ausência de positividade no teste QuickVue® com a carga ou com a cepa viral. O teste QuickVue® mostrou sensibilidade de 90.1%, especificidade 98.9%, valor preditivo positivo 99.3%, valor preditivo negativo de 94.6%, acurácia de 93.2% e índice de concordância de 0.85 em comparação à imunofluorescência. Conclusões: Nosso estudo demonstrou que o teste QuickVue® RSV pode ser efetivo na detecção precoce do vírus sincicial respiratório em amostras de aspirado de nasofaringe e é confiável como uma ferramenta de diagnósticos em pediatria.
Subject(s)
Humans , Male , Female , Child, Preschool , Respiratory Syncytial Viruses/immunology , Respiratory Syncytial Virus Infections/diagnosis , Reverse Transcriptase Polymerase Chain Reaction , Antigens, Viral/analysis , Reagent Kits, Diagnostic , Respiratory Syncytial Viruses/isolation & purification , Respiratory Tract Infections/virology , Brazil , Retrospective Studies , Sensitivity and Specificity , Respiratory Syncytial Virus Infections/virology , Fluorescent Antibody Technique, IndirectABSTRACT
OBJECTIVE: The aim of this study was to evaluate the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA) as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. METHODS: The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. RESULTS: From 313 positive samples by immunofluorescence assays, 282 (90%) were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue® RSV Test and viral load or specific strain. The QuickVue® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. CONCLUSIONS: This study demonstrated that the QuickVue® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics.
Subject(s)
Antigens, Viral/analysis , Respiratory Syncytial Virus Infections/diagnosis , Respiratory Syncytial Viruses/immunology , Reverse Transcriptase Polymerase Chain Reaction , Brazil , Child, Preschool , Female , Fluorescent Antibody Technique, Indirect , Humans , Male , Reagent Kits, Diagnostic , Respiratory Syncytial Virus Infections/virology , Respiratory Syncytial Viruses/isolation & purification , Respiratory Tract Infections/virology , Retrospective Studies , Sensitivity and SpecificityABSTRACT
Objective: The aim of this study was to evaluate the QuickVue (R) RSV Test Kit (QUIDEL Corp, CA, USA) as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. Methods: The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue (R) RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. Results: From 313 positive samples by immunofluorescence assays, 282 (90%) were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays RSV Test and viral load or specific strain. The QuickVue (R) RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement kappa index of 0.85 in comparison to immunofluorescence assay. Conclusions: This study demonstrated that the QuickVue RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics. (C) 2016 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda.
Objetivo Avaliar o teste QuickVue® RSV Test Kit (QUIDEL Corp, CA, EUA) para o diagnóstico rápido do vírus sincicial respiratório em crianças com doença respiratória aguda, comparando-o com a imunofluorescência indireta como padrão ouro. Visto que, no Brasil, testes rápidos para detecção de antígenos para vírus sincicial respiratório não são rotineiramente utilizados como ferramenta de diagnóstico, exceto para Dengue e Influenza. Métodos Um total de 486 amostras de aspirado de nasofaringe de crianças menores de 5 anos com doença respiratória aguda, coletadas entre dezembro de 2013 e agosto de 2014, foram analisadas por imunofluorescência e pelo teste QuickVue®. Amostras com resultados discordantes entre os métodos foram submetidas a PCR em tempo real e sequenciamento. Resultados Das 313 amostras positivas por IFI, 282 foram positivas no teste rápido (90%), 2 amostras foram positivas apenas no teste rápido (0.6%), 33 apenas na imunofluorescência (10.5%) e 171 foram negativas em ambos os métodos. As 35 amostras com resultados discordantes foram testadas por PCR em tempo real, sendo que duas que foram positivas apenas no teste rápido e 5 apenas na imunofluorescência confirmaram-se positivas. Não houve relação entre a ausência de positividade no teste QuickVue® com a carga ou com a cepa viral. O teste QuickVue® mostrou sensibilidade de 90.1%, especificidade 98.9%, valor preditivo positivo 99.3%, valor preditivo negativo de 94.6%, acurácia de 93.2% e índice ? de concordância de 0.85 em comparação à imunofluorescência. Conclusões Nosso estudo demonstrou que o teste QuickVue® RSV pode ser efetivo na detecção precoce do vírus sincicial respiratório em amostras de aspirado de nasofaringe e é confiável como uma ferramenta de diagnósticos em pediatria.
ABSTRACT
OBJECTIVES: Experimental studies on sepsis have demonstrated that ethyl pyruvate is endowed with antioxidant and anti-inflammatory properties. This study aimed to investigate the effects of ethyl pyruvate on leukocyte-endothelial interactions in the mesenteric microcirculation in a live Escherichia coli-induced sepsis model in rats. METHODS: Male Wistar rats were administered an intravenous suspension of E. coli bacteria or were subjected to a sham procedure. Three hours after bacterial infusion, the rats were randomized into the following groups: a control group without treatment, a group treated with lactated Ringer's solution (4 mL/kg, i.v.), and a group treated with lactated Ringer's solution (4 mL/kg, i.v.) plus ethyl pyruvate (50 mg/kg). At 24 h after bacterial infusion, leukocyte-endothelial interactions were investigated using intravital microscopy, and the expression of P-selectin and intercellular adhesion molecule-1 was evaluated via immunohistochemistry. White blood cell and platelet counts were also determined at baseline and 3 h and 24 h after E. coli inoculation. RESULTS: The non-treated and lactated Ringer's solution-treated groups exhibited increases in the numbers of rolling leukocytes (â¼2.5-fold increase), adherent cells (â¼3.0-fold), and migrated cells (â¼3.5-fold) compared with the sham group. In contrast, treatment with Ringer's ethyl pyruvate solution reduced the numbers of rolling, adherent and migrated leukocytes to the levels observed in the sham group. Additionally, the expression of P-selectin and intercellular adhesion molecule-1 was significantly increased on mesenteric microvessels in the non-treated group compared with the sham group (p<0.001). The expression of both adhesion molecules was reduced in the other groups, with ethyl pyruvate being more effective than lactated Ringer's solution. Infusion of bacteria caused significant leukopenia (3 h), followed by leukocytosis with granulocytosis (24 h). There was also an intense and progressive reduction in the number of platelets. However, no differences were observed after treatment with the different solutions. CONCLUSIONS: The presented data suggest that ethyl pyruvate efficiently reduces the inflammatory response in the mesenteric microcirculation in an experimental model of sepsis induced by live E. coli and is associated, at least in part, with down-regulation of P-selectin and intercellular adhesion molecule-1.
Subject(s)
Cell Communication/drug effects , Endothelial Cells/drug effects , Leukocytes/drug effects , Mesenteric Veins/drug effects , Pyruvates/pharmacology , Sepsis/drug therapy , Animals , Cell Communication/physiology , Disease Models, Animal , Endothelial Cells/cytology , Escherichia coli Infections , Leukocytes/cytology , Male , Mesenteric Veins/cytology , Microcirculation , Rats , Rats, WistarABSTRACT
OBJECTIVES: Experimental studies on sepsis have demonstrated that ethyl pyruvate is endowed with antioxidant and anti-inflammatory properties. This study aimed to investigate the effects of ethyl pyruvate on leukocyte-endothelial interactions in the mesenteric microcirculation in a live Escherichia coli-induced sepsis model in rats. METHODS: Male Wistar rats were administered an intravenous suspension of E. coli bacteria or were subjected to a sham procedure. Three hours after bacterial infusion, the rats were randomized into the following groups: a control group without treatment, a group treated with lactated Ringer’s solution (4 mL/kg, i.v.), and a group treated with lactated Ringer’s solution (4 mL/kg, i.v.) plus ethyl pyruvate (50 mg/kg). At 24 h after bacterial infusion, leukocyte-endothelial interactions were investigated using intravital microscopy, and the expression of P-selectin and intercellular adhesion molecule-1 was evaluated via immunohistochemistry. White blood cell and platelet counts were also determined at baseline and 3 h and 24 h after E. coli inoculation. RESULTS: The non-treated and lactated Ringer’s solution-treated groups exhibited increases in the numbers of rolling leukocytes (∼2.5-fold increase), adherent cells (∼3.0-fold), and migrated cells (∼3.5-fold) compared with the sham group. In contrast, treatment with Ringer’s ethyl pyruvate solution reduced the numbers of rolling, adherent and migrated leukocytes to the levels observed in the sham group. Additionally, the expression of P-selectin and intercellular adhesion molecule-1 was significantly increased on mesenteric microvessels in the non-treated group compared with the sham group (p<0.001). The expression of both adhesion molecules was reduced in the other groups, with ethyl pyruvate being more effective than lactated Ringer’s solution. Infusion of bacteria caused significant leukopenia (3 h), followed ...
Subject(s)
Animals , Male , Rats , Cell Communication/drug effects , Endothelial Cells/drug effects , Leukocytes/drug effects , Mesenteric Veins/drug effects , Pyruvates/pharmacology , Sepsis/drug therapy , Cell Communication/physiology , Disease Models, Animal , Escherichia coli Infections , Endothelial Cells/cytology , Leukocytes/cytology , Microcirculation , Mesenteric Veins/cytology , Rats, WistarABSTRACT
INTRODUCTION: Choosing the antimicrobial agent for initial therapy of urinary tract infection (UTI) is usually empirical and should consider the prevalence of uropathogens in different age groups and gender. OBJECTIVE: To establish prevalence rates of uropathogens in community-acquired UTI in relation to age and gender. METHODS: Cross-sectional study conducted in the emergency department (ED) of a general hospital, from January to December, 2010, in patients younger than 15 years old who had clinical suspicion of UTI and collected quantitative urine culture. UTI was defined as urine culture with growth of a single agent > 100.000 colony forming units (cfu)/mL in a midstream collection or ≥ 50.000 cfu/mL in urethral catheterization. RESULTS: There were 63.464 visits to ED. 2577 urine cultures were obtained, of whom 291 were positive for UTI (prevalence = 11.3% of clinical suspicion and 0.46% of visits), 212 cases (72.8%) in females, median age = 2.6 years. The predominant uropathogen was E. coli (76.6%), followed by Proteus mirabilis (10.3%) and Staphylococcus saprophyticus (4.1%). Among infants < 3 months, prevalence rates of E. coli were significantly lower (50% vs 78.4%; OR = 0.276; p = 0.006). Higher prevalences of Staphylococcus saprophyticus occurred among patients > 10 years (24.4% vs 0.4%; OR = 79.265; p < 0.0001). Proteus mirabilis was significantly more prevalent in boys than girls (24.0% vs 5.2%; OR = 5.786; p < 0.001). CONCLUSIONS: E. coli was the most prevalent community-acquired uropathogen. Nevertheless, initial empiric antimicrobial treatment of UTI should consider the significant prevalence of other agents different from E. coli in infants < 3 months, the high prevalence of Staphylococcus saprophyticus in patients > 10 years and Proteus mirabilis in males.
Subject(s)
Bacterial Infections/epidemiology , Urinary Tract Infections/epidemiology , Adolescent , Age Distribution , Age Factors , Bacterial Infections/microbiology , Child , Child, Preschool , Community-Acquired Infections/epidemiology , Community-Acquired Infections/microbiology , Cross-Sectional Studies , Female , Humans , Infant , Infant, Newborn , Male , Prevalence , Retrospective Studies , Sex Distribution , Sex Factors , Urinary Tract Infections/microbiologyABSTRACT
INTRODUÇÃO: A escolha do antimicrobiano para tratamento inicial de infecção de trato urinário (ITU) costuma ser empírica e deve considerar a prevalência dos uropatógenos nas diversas faixas etárias e sexo. OBJETIVO: Avaliar a prevalência de uropatógenos em ITU comunitária e sua relação com idade e sexo. MÉTODOS: Estudo transversal conduzido em pronto socorro (PS) de hospital geral, de janeiro a dezembro, 2010, em pacientes menores de 15 anos com suspeita clínica de ITU, que colheram urocultura quantitativa. Definida ITU como urocultura com crescimento de agente único > 100.000 unidades formadoras de colônia (ufc)/mL na coleta por jato médio ou > 50.000 ufc/mL na coleta por sondagem vesical. RESULTADOS: Ocorreram 63.464 atendimentos no PS. Foram obtidas 2.577 uroculturas; destas, 291 foram positivas para ITU (prevalência = 11,3% das suspeitas clínicas e 0,46% dos atendimentos); 212 casos (72,8%) em meninas, mediana de idade = 2,6 anos. O uropatógeno predominante foi E.coli (76,6%), seguido por Proteus mirabilis (10,3%) e Staphylococcus saprophyticus (4,1%). Em lactentes < 3 meses, a prevalência de E.coli foi significativamente menor (50% x 78,4%; OR = 0,276; p = 0,006). Maior prevalência de Staphylococcus saprophyticus ocorreu em pacientes > 10 anos (24,4% x 0,4%; OR = 79,265; p < 0,0001). Proteus mirabilis foi significativamente mais prevalente em meninos (24,0% x 5,2%; OR = 5,786; p < 0,001). CONCLUSÕES: E. coli foi o uropatógeno mais prevalente das ITU comunitárias. Entretanto, na escolha do antimicrobiano empírico inicial, deve-se levar em consideração a prevalência significativa de outros agentes diferentes de E. coli em lactentes < 3 meses, a alta prevalência de Staphylococcus saprophyticus em pacientes > 10 anos e de Proteus mirabilis em meninos.
INTRODUCTION: Choosing the antimicrobial agent for initial therapy of urinary tract infection (UTI) is usually empirical and should consider the prevalence of uropathogens in different age groups and gender. OBJECTIVE: To establish prevalence rates of uropathogens in community-acquired UTI in relation to age and gender. METHODS: Crosssectional study conducted in the emergency department (ED) of a general hospital, from January to December, 2010, in patients younger than 15 years old who had clinical suspicion of UTI and collected quantitative urine culture. UTI was defined as urine culture with growth of a single agent > 100.000 colony forming units (cfu)/mL in a midstream collection or > 50.000 cfu/mL in urethral catheterization. RESULTS: There were 63.464 visits to ED. 2577 urine cultures were obtained, of whom 291 were positive for UTI (prevalence = 11.3% of clinical suspicion and 0.46% of visits), 212 cases (72.8%) in females, median age = 2.6 years. The predominant uropathogen was E. coli (76.6%), followed by Proteus mirabilis (10.3%) and Staphylococcus saprophyticus (4.1%). Among infants < 3 months, prevalence rates of E. coli were significantly lower (50% vs 78.4%; OR = 0.276; p = 0.006). Higher prevalences of Staphylococcus saprophyticus occurred among patients > 10 years (24.4% vs 0.4%; OR = 79.265; p < 0.0001). Proteus mirabilis was significantly more prevalent in boys than girls (24.0% vs 5.2%; OR = 5.786; p < 0.001). CONCLUSIONS: E. coli was the most prevalent community-acquired uropathogen. Nevertheless, initial empiric antimicrobial treatment of UTI should consider the significant prevalence of other agents different from E. coli in infants < 3 months, the high prevalence of Staphylococcus saprophyticus in patients > 10 years and Proteus mirabilis in males.
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Bacterial Infections/epidemiology , Urinary Tract Infections/epidemiology , Age Distribution , Age Factors , Bacterial Infections/microbiology , Cross-Sectional Studies , Community-Acquired Infections/epidemiology , Community-Acquired Infections/microbiology , Prevalence , Retrospective Studies , Sex Distribution , Sex Factors , Urinary Tract Infections/microbiologyABSTRACT
OBJECTIVE: To evaluate the impact of the routine use of rapid antigen detection test in the diagnosis and treatment of acute pharyngotonsillitis in children. METHODS: This is a prospective and observational study, with a protocol compliance design established at the Emergency Unit of the University Hospital of Universidade de São Paulo for the care of children and adolescents diagnosed with acute pharyngitis. RESULTS: 650 children and adolescents were enrolled. Based on clinical findings, antibiotics would be prescribed for 389 patients (59.8%); using the rapid antigen detection test, they were prescribed for 286 patients (44.0%). Among the 261 children who would not have received antibiotics based on the clinical evaluation, 111 (42.5%) had positive rapid antigen detection test. The diagnosis based only on clinical evaluation showed 61.1% sensitivity, 47.7% specificity, 44.9% positive predictive value, and 57.5% negative predictive value. CONCLUSIONS: The clinical diagnosis of streptococcal pharyngotonsillitis had low sensitivity and specificity. The routine use of rapid antigen detection test led to the reduction of antibiotic use and the identification of a risk group for complications of streptococcal infection, since 42.5% positive rapid antigen detection test patients would not have received antibiotics based only on clinical diagnosis.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Antigens, Bacterial/analysis , Pharyngitis/diagnosis , Pharyngitis/drug therapy , Streptococcal Infections/diagnosis , Streptococcal Infections/therapy , Streptococcus pyogenes/immunology , Tonsillitis/diagnosis , Tonsillitis/drug therapy , Acute Disease , Adolescent , Child , Child, Preschool , Emergency Service, Hospital , Female , Humans , Male , Pharyngitis/complications , Prospective Studies , Respiratory Tract Infections , Time Factors , Tonsillitis/complicationsABSTRACT
OBJETIVO: Avaliar o impacto da realização rotineira da prova rápida para pesquisa de estreptococo do grupo A no diagnóstico e tratamento da faringotonsilite aguda em crianças. MÉTODOS: Estudo prospectivo e observacional que contou com a utilização de protocolo de pesquisa estabelecido na Unidade de Emergência do Hospital Universitário da Universidade de São Paulo para o atendimento de crianças e adolescentes com faringotonsilite aguda. RESULTADOS: Com base na avaliação clínica, dos 650 pacientes estudados, antimicrobianos seriam prescritos para 389 indivíduos (59,8%) e, com o uso da pesquisa de estreptococo do grupo A, foram prescritos em 286 pacientes (44,0%). Das 261 crianças que não receberiam antibiótico pelo quadro clínico, 111 (42,5%) tiveram pesquisa de estreptococo do grupo A positiva. O diagnóstico baseado no quadro clínico apresentou sensibilidade de 61,1%, especificidade de 47,7%, valor preditivo positivo de 44,9% e valor preditivo negativo de 57,5%. CONCLUSÕES: O diagnóstico clínico da faringotonsilite estreptocócica mostrou baixa sensibilidade e especificidade. O uso rotineiro da prova rápida para pesquisa de estreptococo permitiu a redução do uso de antibióticos e a identificação de um grupo de risco para as complicações da infecção estreptocócica, pois 42,5% dos pacientes com prova rápida positiva não receberiam antibióticos, se levado em consideração apenas o diagnóstico clínico.
OBJECTIVE: To evaluate the impact of the routine use of rapid antigen detection test in the diagnosis and treatment of acute pharyngotonsillitis in children. METHODS: This is a prospective and observational study, with a protocol compliance design established at the Emergency Unit of the University Hospital of Universidade de São Paulo for the care of children and adolescents diagnosed with acute pharyngitis. RESULTS: 650 children and adolescents were enrolled. Based on clinical findings, antibiotics would be prescribed for 389 patients (59.8%); using the rapid antigen detection test, they were prescribed for 286 patients (44.0%). Among the 261 children who would not have received antibiotics based on the clinical evaluation, 111 (42.5%) had positive rapid antigen detection test. The diagnosis based only on clinical evaluation showed 61.1% sensitivity, 47.7% specificity, 44.9% positive predictive value, and 57.5% negative predictive value. CONCLUSIONS: The clinical diagnosis of streptococcal pharyngotonsillitis had low sensitivity and specificity. The routine use of rapid antigen detection test led to the reduction of antibiotic use and the identification of a risk group for complications of streptococcal infection, since 42.5% positive rapid antigen detection test patients would not have received antibiotics based only on clinical diagnosis.
OBJETIVO: Evaluar el impacto de la realización de rutina de la prueba rápida para investigación de estreptococos del grupo A en el diagnóstico y tratamiento de la faringotonsilitis aguda en niños. MÉTODOS: Estudio prospectivo y observacional que contó con el uso de protocolo de investigación establecido en la Unidad de Emergencia del Hospital Universitario de la USP para la atención a niños y adolescentes con faringotonsilitis aguda. RESULTADOS: Con base en la evaluación crítica, de los 650 pacientes estudiados, antimicrobianos serían prescritos a 389 individuos (59,8%) y, con el uso de la investigación de estreptococos del grupo A se los prescribieron a 286 pacientes (44,0%). De los 261 niños que no recibirían antibióticos por el cuadro clínico, 111 (42,5%) tuvieron investigación de estreptococos del grupo A positiva. El diagnóstico basado en el cuadro clínico presentó sensibilidad del 61,1%, especificidad del 47,7%, valor predictivo positivo del 44,9% y valor predictivo negativo del 57,5%, CONCLUSIONES: En este estudio, el diagnóstico clínico de la faringotonsilitis estreptocócica mostró baja sensibilidad y especificidad. El uso de rutina de la prueba rápida para investigación de estreptococos permitió la reducción del uso de antibióticos y la identificación de un grupo de riesgo para las complicaciones de la infección estreptocócica, pues el 42,5% de los pacientes con prueba rápida positiva no recibirían antibióticos si se llevara en consideración solamente el diagnóstico clínico.
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Anti-Bacterial Agents/therapeutic use , Antigens, Bacterial/analysis , Pharyngitis/diagnosis , Pharyngitis/drug therapy , Streptococcal Infections/diagnosis , Streptococcal Infections/therapy , Streptococcus pyogenes/immunology , Tonsillitis/diagnosis , Tonsillitis/drug therapy , Acute Disease , Emergency Service, Hospital , Pharyngitis/complications , Prospective Studies , Respiratory Tract Infections , Time Factors , Tonsillitis/complicationsABSTRACT
The innate and adaptive immune responses of dendritic cells (DCs) to enteroinvasive Escherichia coli (EIEC) infection were compared with DC responses to Shigella flexneri infection. EIEC triggered DCs to produce interleukin (IL)-10, IL-12 and tumour necrosis factor (TNF)-α, whereas S. flexneri induced only the production of TNF-α. Unlike S. flexneri, EIEC strongly increased the expression of toll like receptor (TLR)-4 and TLR-5 in DCs and diminished the expression of co-stimulatory molecules that may cooperate to inhibit CD4(+) T-lymphocyte proliferation. The inflammation elicited by EIEC seems to be related to innate immunity both because of the aforementioned results and because only EIEC were able to stimulate DC transmigration across polarised Caco-2 cell monolayers, a mechanism likely to be associated with the secretion of CC chemokine ligands (CCL)20 and TNF-α. Understanding intestinal DC biology is critical to unravelling the infection strategies of EIEC and may aid in the design of treatments for infectious diseases.
Subject(s)
Dendritic Cells/immunology , Escherichia coli/immunology , Immunity, Innate/immunology , Immunity, Mucosal/immunology , Intestinal Mucosa/microbiology , Shigella flexneri/immunology , Animals , CD4 Lymphocyte Count , Cell Proliferation , Humans , Interleukin-10/biosynthesis , Interleukin-12/biosynthesis , Intestinal Mucosa/cytology , Intestinal Mucosa/immunology , Murinae , Toll-Like Receptor 4/immunology , Toll-Like Receptor 5/immunology , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
The innate and adaptive immune responses of dendritic cells (DCs) to enteroinvasive Escherichia coli (EIEC) infection were compared with DC responses to Shigella flexneri infection. EIEC triggered DCs to produce interleukin (IL)-10, IL-12 and tumour necrosis factor (TNF)-α, whereas S. flexneri induced only the production of TNF-α. Unlike S. flexneri, EIEC strongly increased the expression of toll like receptor (TLR)-4 and TLR-5 in DCs and diminished the expression of co-stimulatory molecules that may cooperate to inhibit CD4+ T-lymphocyte proliferation. The inflammation elicited by EIEC seems to be related to innate immunity both because of the aforementioned results and because only EIEC were able to stimulate DC transmigration across polarised Caco-2 cell monolayers, a mechanism likely to be associated with the secretion of CC chemokine ligands (CCL)20 and TNF-α. Understanding intestinal DC biology is critical to unravelling the infection strategies of EIEC and may aid in the design of treatments for infectious diseases.
Subject(s)
Animals , Humans , Dendritic Cells/immunology , Escherichia coli/immunology , Immunity, Innate/immunology , Immunity, Mucosal/immunology , Intestinal Mucosa/microbiology , Shigella flexneri/immunology , Cell Proliferation , /biosynthesis , /biosynthesis , Intestinal Mucosa/cytology , Intestinal Mucosa/immunology , Murinae , /immunology , /immunology , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
This study evaluated the polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) analysis of fliC for typing flagella antigen (H) of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) strains isolated from different animals. The molecular typing of the H type was efficient in the determination of 93 (85%) strains. Two nonmotile (H-) E. coli strains showed a PCR-RFLP electrophoretic profile that did not match known H type patterns. The fliC nucleotide sequence of strains B2N and 4a revealed a nucleotide substitution at the restriction site and a nucleotide insertion that generated a stop codon, respectively. The results of this study showed that PCR-RFLP analysis of fliC is faster, less laborious and as efficient for the determination of H type E. coli isolated from animals, compared to serotyping and that it is useful in determining H type in nonmotile strains and strains expressing non-reactive H antigens. Moreover, the fliC sequence of strain B2N suggests that we could have found a new flagellin antigen type.
Subject(s)
Enteropathogenic Escherichia coli/classification , Escherichia coli Proteins/genetics , Molecular Typing/veterinary , Polymorphism, Restriction Fragment Length , Shiga-Toxigenic Escherichia coli/classification , Animals , Antigens, Bacterial/genetics , Base Sequence , Brazil , Cattle , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Dogs , Enteropathogenic Escherichia coli/genetics , Enteropathogenic Escherichia coli/isolation & purification , Flagellin , Haplorhini , Molecular Sequence Data , Molecular Typing/methods , Sequence Analysis, DNA/veterinary , Sheep , Shiga-Toxigenic Escherichia coli/genetics , Shiga-Toxigenic Escherichia coli/isolation & purificationABSTRACT
OBJETIVO: Descrever a prevalência dos principais agentes etiológicos de infecção urinária comunitária em crianças menores de 15 anos e analisar o perfil de sensibilidade antimicrobiana do principal agente, Escherichia coli. MÉTODOS: Estudo retrospectivo conduzido a partir de uroculturas com crescimento de agente único, com unidades formadoras de colônia maior ou igual a 10(5)/mL. A população selecionada foi atendida no Pronto Atendimento de Pediatria do Hospital Universitário da Universidade de São Paulo entre janeiro de 2006 e dezembro de 2007. Para analisar o perfil de sensibilidade de E. coli, foram excluídos casos com infecção urinária de repetição e/ou portadores de uropatias. RESULTADOS: Dos 176 casos de infecção urinária, 133 (76 por cento) ocorreram no sexo feminino e 43 (24 por cento) no masculino. A faixa etária de maior prevalência das infecções urinárias foi a de menores de dois anos, com 84 casos (48 por cento). Escherichia coli foi o principal agente isolado (122 culturas - 69 por cento do total). Foram encontrados altos índices de sensibilidade antimicrobiana de E. coli às cefalosporinas de segunda e terceira geração (cefuroxima e ceftriaxona, 100 por cento), aos aminoglicosídeos (amicacina, 100 por cento; gentamicina, 96,4 por cento), ao ácido nalidíxico (97,3 por cento), à nitrofurantoina (98,2 por cento) e às quinolonas (ciprofloxacina e norfloxacina, 98,2 por cento); sensibilidade moderada à cefalosporina de primeira geração (cefalotina, 78,4 por cento); baixa sensibilidade à ampicilina (38,7 por cento) e ao sufametoxazol-trimetoprim (55 por cento). CONCLUSÕES: E. coli continua sendo a bactéria mais prevalente das infecções urinárias comunitárias. Conhecendo a baixa sensibilidade in vitro dessa bactéria à sulfametoxazol-trimetoprim e à ampicilina, recomenda-se que tais drogas não sejam eleitas para a terapêutica inicial.
OBJECTIVE: To describe the prevalence of the most frequent etiological agents of community acquired urinary tract infection in children under the age of 15 years, as well as to analyse the antimicrobial susceptibility patterns of the most prevalent pathogen, Escherichia coli. METHODS: Retrospective study conducted from urine cultures of a single bacterial species, at a concentration > 10(5) colonies forming units/mL. The children included in the study were treated in the Pediatric Emergency Care Unit of the University Hospital of the University of Sao Paulo from January, 2006 to December, 2007. In order to analyze Escherichia coli antimicrobial susceptibility patterns, cases with recurrent urinary tract infection and/or carriers of uropathy were excluded. RESULTS: Among 176 urinary tract infection patients, 133 (76 percent) were females. Children less than two years old presented the highest prevalence of urinary tract infection (84 cases; 48 percent). E. coli was the main agent (122 cultures - 69 percent of the total). There were high rates of E. coli antimicrobial susceptibility to second and third-generation cephalosporins (cefuroxime and ceftriaxone, 100 percent), aminoglycosides (amikacin, 100 percent, gentamicin, 96.4 percent), nalidixic acid (97.3 percent), nitrofurantoin (98.2 percent) and quinolones (ciprofloxacin and norfloxacin, 98.2 percent); moderate susceptibility to first-generation cephalosporin (cephalothin, 78.4 percent); low susceptibility to ampicillin (38.7 percent) and sufamethoxazole-trimethoprim (55 percent). CONCLUSIONS: E. coli remains the most prevalent bacterial pathogen of community acquired urinary tract infection. Given the low in vitro susceptibility of these bacteria to sulfamethoxazole-trimethoprim and ampicillin, these drugs should not be elected as the initial choice for treatment.
Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Escherichia coli , Urinary Tract Infections/etiology , Microbial Sensitivity TestsABSTRACT
In a study conducted in João Pessoa, northeast of Brazil, 2344 Escherichia coli isolated from 290 infants with diarrhea and 290 healthy matched controls were analyzed for virulence traits. Enteroaggregative E. coli (EAEC) was the most prevalent pathogen associated to acute diarrhea. Based on the results of colony blot hybridization, serotyping, and HEp-2 cell adherence assays, strains were separated in categories as typical enteropathogenic E. coli (EPEC) (1.7%), atypical EPEC (a-EPEC) (9.3%), EAEC (25%), enterotoxigenic E. coli (10%), and enteroinvasive E. coli (EIEC) (1.4%). No enterohemorrhagic E. coli strains were isolated. Other enteropathogens were found, including Salmonella (7.9%), Shigella spp. (4.1%), thermophilic Campylobacter spp. (2.4%), Giardia lamblia (9.3%), and Entamoeba histolytica (5.8%). All enteropathogens were associated with diarrhea (P < 0.01). However, the association was lower for EPEC and EIEC (P < 0.03). Different pathogens associated with diarrhea may have been changing in Brazil where EAEC and a-EPEC seem to be the most prevalent pathogens among them.
Subject(s)
Diarrhea, Infantile/microbiology , Enteropathogenic Escherichia coli/classification , Escherichia coli Infections/microbiology , Brazil/epidemiology , Case-Control Studies , Cell Adhesion , Cell Line, Tumor , Chi-Square Distribution , Diarrhea, Infantile/epidemiology , Enteropathogenic Escherichia coli/genetics , Escherichia coli/classification , Escherichia coli/genetics , Escherichia coli Infections/epidemiology , Feces/microbiology , Gram-Negative Bacteria/classification , Gram-Negative Bacteria/genetics , Gram-Negative Bacterial Infections/epidemiology , Gram-Negative Bacterial Infections/microbiology , Humans , Infant , Prevalence , Serotyping , VirulenceABSTRACT
Important features of the enteroinvasive Escherichia coli (EIEC) phenotype and gene expression likely to confer EIEC with a lower ability to cause disease than Shigella flexneri were described here for the first time. To confirm the lower pathogenicity of EIEC, we have analyzed the keratoconjunctivitis developed in guinea-pigs with EIEC or S. flexneri. Shigella flexneri induced a more pronounced proinflammatory response, whereas EIEC induced a mild form of the disease. EIEC showed a significantly less efficient cell-to-cell Caco-2 dissemination when compared with S. flexneri. Plaques formed by EIEC during intercellular spreading were four times smaller than those formed by S. flexneri. At the molecular level, the lower expression of virulence genes by EIEC during infection of Caco-2 cells highlighted the importance of effective gene transcription for bacterial pathogenicity.
Subject(s)
Bacterial Proteins/biosynthesis , Escherichia coli/pathogenicity , Gene Expression , Shigella flexneri/pathogenicity , Virulence Factors/biosynthesis , Animals , Bacterial Proteins/genetics , Caco-2 Cells , Colony Count, Microbial , Conjunctiva/microbiology , Conjunctiva/pathology , Disease Models, Animal , Epithelial Cells/microbiology , Gene Expression Profiling , Guinea Pigs , Humans , Keratoconjunctivitis/microbiology , Virulence , Virulence Factors/geneticsABSTRACT
PURPOSE: Bacterial translocation has been shown to occur in critically ill patients after extensive trauma, shock, sepsis, or thermal injury. The present study investigates mesenteric microcirculatory dysfunctions, the bacterial translocation phenomenon, and hemodynamic/metabolic disturbances in a rat model of intestinal obstruction and ischemia. METHODS: Anesthetized (pentobarbital 50 mg/kg, i.p.) male Wistar rats (250-350 g) were submitted to intestinal obstruction or laparotomy without intestinal obstruction (Sham) and were evaluated 24 hours later. Bacterial translocation was assessed by bacterial culture of the mesenteric lymph nodes (MLN), liver, spleen, and blood. Leukocyte-endothelial interactions in the mesenteric microcirculation were assessed by intravital microscopy, and P-selectin and intercellular adhesion molecule (ICAM)-1 expressions were quantified by immunohistochemistry. Hematocrit, blood gases, lactate, glucose, white blood cells, serum urea, creatinine, bilirubin, and hepatic enzymes were measured. RESULTS: About 86% of intestinal obstruction rats presented positive cultures for E. coli in samples of the mesenteric lymph nodes, liver, and spleen, and 57% had positive hemocultures. In comparison to the Sham rats, intestinal obstruction induced neutrophilia and increased the number of rolling (approximately 2-fold), adherent (approximately 5-fold), and migrated leukocytes (approximately 11-fold); this increase was accompanied by an increased expression of P-selectin (approximately 2-fold) and intercellular adhesion molecule-1 (approximately 2-fold) in the mesenteric microcirculation. Intestinal obstruction rats exhibited decreased PaCO2, alkalosis, hyperlactatemia, and hyperglycemia, and increased blood potassium, hepatic enzyme activity, serum urea, creatinine, and bilirubin. A high mortality rate was observed after intestinal obstruction (83% at 72 h vs. 0% in Sham rats). CONCLUSION: Intestinal obstruction and ischemia in rats is a relevant model for the in vivo study of mesenteric microcirculatory dysfunction and the occurrence of bacterial translocation. This model parallels the events implicated in multiple organ dysfunction (MOD) and death.
Subject(s)
Bacterial Translocation/physiology , Escherichia coli/physiology , Intestinal Obstruction/physiopathology , Intestine, Small/blood supply , Ischemia/physiopathology , Microcirculation/physiology , Animals , Biomarkers/blood , Disease Models, Animal , Immunohistochemistry , Intestinal Obstruction/blood , Intestinal Obstruction/microbiology , Intestine, Small/microbiology , Intestine, Small/physiopathology , Male , Multiple Organ Failure/physiopathology , Rats , Rats, WistarABSTRACT
Introdução: Faringoamidalite é queixa frequente nos pronto-socorros pediátricos, com presença do Streptococcus pyogenes em 20 por cento dos casos. Os sinais e sintomas clínicos são insuficientes para diferenciar quadros virais dos bacterianos. Os métodos microbiológicos, teste rápido(TR) e cultura orofaringe(CO) são indispensáveis para diagnóstico e redução de antibióticos...
Introductions: Pharyngitis is a common complaint in the emergency room for children, with the presence of Streptococcus pyogenes in 20 per cent of the cases. The clinical signs and symptoms are insufficient to differentiate viral from bacterial infections. The microbiological methods, rapid test (RT) and oropharynx cultura (OC), are essential for diagnosis and for reducing the use of antibiotics...
Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Pharyngitis/diagnosis , Pharyngitis/microbiology , Streptococcal Infections/diagnosis , Streptococcal Infections/microbiology , Streptococcus/pathogenicity , Tonsillitis/diagnosis , Tonsillitis/microbiology , Culture Media , Prospective Studies , Sensitivity and Specificity , Streptococcus/isolation & purificationABSTRACT
PRUPOSE: Bacterial translocation has been shown to occur in critically ill patients after extensive trauma, shock, sepsis, or thermal injury. The present study investigates mesenteric microcirculatory dysfunctions, the bacterial translocation phenomenon, and hemodynamic/metabolic disturbances in a rat model of intestinal obstruction and ischemia. METHODS: Anesthetized (pentobarbital 50 mg/kg, i.p.) male Wistar rats (250-350 g) were submitted to intestinal obstruction or laparotomy without intestinal obstruction (Sham) and were evaluated 24 hours later. Bacterial translocation was assessed by bacterial culture of the mesenteric lymph nodes (MLN), liver, spleen, and blood. Leukocyte-endothelial interactions in the mesenteric microcirculation were assessed by intravital microscopy, and P-selectin and intercellular adhesion molecule (ICAM)-1 expressions were quantified by immunohistochemistry. Hematocrit, blood gases, lactate, glucose, white blood cells, serum urea, creatinine, bilirubin, and hepatic enzymes were measured. RESULTS: About 86 percent of intestinal obstruction rats presented positive cultures for E. coli in samples of the mesenteric lymph nodes, liver, and spleen, and 57 percent had positive hemocultures. In comparison to the Sham rats, intestinal obstruction induced neutrophilia and increased the number of rolling (~2-fold), adherent (~5-fold), and migrated leukocytes (~11-fold); this increase was accompanied by an increased expression of P-selectin (~2-fold) and intercellular adhesion molecule-1 (~2-fold) in the mesenteric microcirculation. Intestinal obstruction rats exhibited decreased PaCO2, alkalosis, hyperlactatemia, and hyperglycemia, and increased blood potassium, hepatic enzyme activity, serum urea, creatinine, and bilirubin. A high mortality rate was observed after intestinal obstruction (83 percent at 72 h vs. 0 percent in Sham rats). CONCLUSION: Intestinal obstruction and ischemia in rats is a relevant model for ...