Assessing the impact of the Gamma variant on COVID-19 patient admissions in a southern Brazilian tertiary hospital-A comparison of dual pandemic phases.

Since the first case of COVID-19, Brazil has undergone infection waves with distinct characteristics. The description of new variants has alerted the emergence of more contagious or virulent viruses. The variant of concern Gamma emerged in Brazil and caused an epidemic wave, but its spread outside the country was limited. We report the clinical-epidemiological profile of hospitalized patients with COVID-19 by comparing two periods. A retrospective cohort study was performed. The primary outcome was to assess individuals with COVID-19 admitted in wards and intensive care units at the academic hospital of the Federal University of Parana (CHC-UFPR) between March 2020 and July 2021, correlating demographic, clinical-epidemiologic, and survival data with the most prevalent viral variant found in each period. We used Kaplan-Meier analysis to estimate the probability of survival and ROC curves to evaluate laboratory tests to find a cutoff point for poor outcomes. Data from 2,887 individuals were analyzed, 1,495 and 1,392 from the first and second periods, respectively. Hospitalization predominated among males in both periods, and the median age was significantly lower in the second one. The frequency of comorbidities was similar. Various demographic factors, clinical assessments, and laboratory tests were examined in relation to greater severity. When comparing the two periods, we observed predominance of the Wild virus during the first wave and the Gamma variant during the second, with no significant difference in outcomes. The findings suggest that despite the association of many factors with increased severity, the temporal variation between the two periods did not result in a notable divergence in the measured outcomes. The COVID-19 pandemic has lasted for a long time, with periods marked by peaks of cases, often caused by the emergence of viral variants, resulting in higher infection rates and rapid dissemination but, for variant Gamma, no apparent greater virulence.

Respiratory syncytial virus: host genetic susceptibility and factors associated with disease severity in a cohort of pediatric patients.

BACKGROUND: Respiratory syncytial virus (RSV) infections are the leading cause of hospitalization in young children. We assessed the epidemiology, severity, clinical characteristics, molecular profile and genetic factors of RSV infections compared to acute respiratory illness (ARI) caused by other respiratory viruses. METHODS: Prospective cohort study was conducted from 2017 to 2018 with children under 2 years old hospitalized with ARI. Detection of respiratory viruses was carried out using RT-PCR. RSVs were genotyped via nucleotide sequencing, and host interleukin 28B (IL28B) single nucleotide polymorphisms (SNPs) were determined using SNP TaqMan® Genotyping Assays. RESULTS: A total of 468 children were included; 288 (61.5%) had an infection by a single virus: 202 (70.1%) cases by RSV followed by rhinovirus 36 (12.5%) and influenza 16 (5.6%). Of the RSV cases, 36% were genotyped with a higher prevalence of RSV B (62.1%). The RSV group presented median age of 2.7 months (1.6-6.8), higher frequency in: intensive care unit admission (p = 0.004), mechanical ventilation use (p = 0.018), wheezing (p < 0.001), antimicrobial use (p < 0.001) and low oxygen saturation (p < 0.001). Prematurity (27.2%) was the most frequent comorbidity. RSV patients without comorbidities demonstrated a higher frequency in the combination of IL28B rs12979860 CT/IL28B rs8099917 TG and IL28B rs12979860 TT/IL28B rs8099917 TT genotypes. Viral coinfection was detected in 27 (5.7%) children, with the most frequent being RSV and rhinovirus (14.2%). CONCLUSIONS: This study highlighted the burden of RSV infection in children under 2 years of age, without comorbidities, with a higher need for pediatric ICU admission. Some IL28B allele combinations had a significant association with RSV frequency of infections.

Rapid Serological Tests for SARS-CoV-2: Diagnostic Performance of 4 Commercial Assays.

OBJECTIVE: To assess the diagnostic performance of lateral flow immunochromatographic assays (LFAs) of 4 different manufacturers to identify SARS-CoV-2 antibodies (IgM, IgG, or total), comparing them with the nucleic acid amplification test (NAAT) or the clinical defined test (definite or probable SARS-CoV-2 infection, respectively). METHODS: One hundred nineteen serum samples were randomly selected by convenience and distributed in the following groups: (1) group with SARS-CoV-2 infection (n = 82; RT-qPCR positive [definite, n = 70] and probable [n = 12]); (2) other diseases (n = 27; other viruses identified [n = 8] and SARS of other etiologies [n = 19]); and (3) healthy control group (n = 10). LFAs of 4 manufacturers were compared: MedTest Coronavirus (COVID-19) IgG/IgM (MedLevensohn, Brazil); COVID-19 IgG/IgM ECO Test (Ecodiagnóstica, Brazil); Camtech COVID-19 IgM/IgG Rapid Test Kit (Camtech Diagnostics Pte Ltd, Singapore); and 1-Step COVID-19 Test for total antibodies (Guangzhou Wondfo Biotech Co., China). RESULTS: The 4 tests studied showed high diagnostic performance characteristics for the diagnoses of definite or probable SARS-CoV-2 infection. The best measures were for the Wondfo test: sensitivity (86.59%; 95% CI: 77.26-93.11%), specificity (100%; 90.51-100%), DOR (257; 60-1,008), LR+ (33.43; 4.82-231.85), LR- (0.13; 0.08-0.23), accuracy (90.76%; 84.06-95.29%), and Matthews correlation coefficient (MCC) 0.82. Although considering only the probable SARS-CoV-2 infection (PCR-) cases, all the kits studied showed limited values. CONCLUSION: Our data demonstrate the excellent performance of LFA for the diagnoses of definite or probable SARS-CoV-2 infection. There was substantial heterogeneity in sensitivities of IgM and IgG antibodies among the different kits. LFA tests cannot replace molecular diagnostics but should be used as an additional screening tool.

Procalcitonin, C-Reactive Protein, Albumin, and Blood Cultures as Early Markers of Sepsis Diagnosis or Predictors of Outcome: A Prospective Analysis.

PURPOSE: Sepsis is a condition with high mortality rates and its diagnosis remains a challenge. We assessed epidemiological, clinical data, multiple biomarker profiles, and blood culture with respect to sepsis diagnosis and predictors of outcome. METHODS: In total, 183 patients who were suspected of having sepsis and underwent blood culture collection were followed up for 7 days. Sepsis-related Organ Failure Assessment (SOFA) and Acute Physiology and Chronic Health Evaluation (APACHE) II scores were calculated daily; biomarkers and blood culture test results were evaluated. RESULTS: In total, 78 (43%) had sepsis, 50 (27%) had septic shock, and 55 (30%) had no sepsis. Blood culture was positive in 28% and 42% of the sepsis and septic shock groups, respectively (P < .001). Regarding clinical profiles and biomarker values, there were no differences between the sepsis and non-sepsis groups, but significant differences were observed in the septic shock group. Multivariate logistic regression models revealed that age, serum albumin level, APACHE II, and SOFA 1st day scores were the independent variables for death. CONCLUSIONS: The challenge in the diagnosis of sepsis continues as clinical and laboratory differences found between the groups were due to septic shock. Older aged patients with lower albumin levels and higher APACHE II and SOFA 1st day scores have a greater probability of mortality.

Global Influenza Hospital-based Surveillance Network (GIHSN): results of surveillance of influenza and other respiratory viruses in hospitalised patients in Brazil, 2015.

BACKGROUND: Influenza-like illness occurs annually worldwide, with peak timing and severity varying seasonally, resulting in significant annual mortality. OBJECTIVES: There were three objectives: (1) to describe the epidemiological and clinical features of hospitalised patients with severe acute respiratory infection caused by influenza and other respiratory viruses (ORVs); (2) to report the influenza seasonality in the region and (3) to correlate findings of influenza circulation and immunisation time in Brazil. PATIENTS/METHODS: This study took place in three Brazilian hospitals located in cities with different climatic conditions (Curitiba (south), Rio de Janeiro (south-east) and Fortaleza (north-east)). Patients presenting with an acute process with indication for admission consisting of a predefined set of conditions potentially associated with recent influenza infection were enrolled. RESULTS: We screened 1666 patients, with 595 meeting the inclusion criteria. Influenza viruses and ORVs were detected in 6.5% and 59% of patients, respectively. Influenza-positive cases fell into the severe spectrum as compared with those with ORVs (30% vs 11%), but without any difference in mortality rates. Epidemiological results revealed variations in the peak time of influenza infections between north-east (Fortaleza) and south (Curitiba) Brazil, basically following the rain period of each region. In north-east Brazil, viral circulation was prevalent in the first 4 months of the year, indicating that the vaccination campaign occurred in a postseasonal period, possibly explaining the low effectiveness. CONCLUSIONS: The active-surveillance model is a valuable tool for investigating respiratory virus impact on hospitalised patients, with influenza-infection monitoring enabling implementation of adequate preventive measures.

Molecular epidemiology of influenza B virus and implications in immunization strategy, Southern Brazil.

Epidemiological indicators have shown the substantial impact of influenza B (Flu B) on the development of severe acute respiratory infection (SARI) and on mortality rates. In Brazil, the trivalent vaccine, composed of only one Flu B lineage is available. We investigated Flu B infections in clinical samples collected by the epidemiological surveillance service of Paraná State, Brazil, from 2013 to 2016. The Flu B lineages Yamagata- (B/Yam) and Victoria-like (B/Vic) were identified using the qRT-PCR assay, and notification forms were reviewed. Among 379 Flu B positive samples evaluated, 370 (98%) were characterized as B/Yam or B/Vic lineages. Both co-circulated with a frequency of 47% and 53%, respectively. B/Yam infected equally both genders, while B/Vic was more frequent in females (71%). The median age of patients infected by B/Vic (23y; 11-35) was lower than that of patients infected by B/Yam (32y; 12-50). Mismatch between the vaccine and the circulating strain was observed in the 2013 season, with a high number of SARI cases. B/Vic lineage was associated with a larger number of SARI cases (62%), while B/Yam with influenza-like illness (ILI) (61%). Differences were observed in the strains circulating in separate regions of Paraná State. B/Vic was prevalent in the northwestern (67%) and B/Yam in the southeastern region (60%). The unpredictability of Flu B lineage circulation causes a substantial increase in severe disease during epidemics in a vaccine mismatch season. In addition, the differences in the epidemiological profile of the target population of Flu B infections in relation to other respiratory viruses, as well as among the B/Vic and B/Yam lineages may also be associated to an increase in disease burden. These findings have direct consequences on vaccination strategies. Therefore, further molecular epidemiology studies of Flu B in Brazil are required to corroborate these primary results.

Comparison of DNA extraction methods used to detect bacterial and yeast DNA from spiked whole blood by real-time PCR.

Sepsis is the leading cause of death in intensive care units (ICUs) worldwide and its diagnosis remains a challenge. Blood culturing is the gold standard technique for blood stream infection (BSI) identification. Molecular tests to detect pathogens in whole blood enable early use of antimicrobials and affect clinical outcomes. Here, using real-time PCR, we evaluated DNA extraction using seven manual and three automated commercially available systems with whole blood samples artificially contaminated with Escherichia coli, Staphylococcus aureus, and Candida albicans, microorganisms commonly associated with BSI. Overall, the commercial kits evaluated presented several technical limitations including long turnaround time and low DNA yield and purity. The performance of the kits was comparable for detection of high microorganism loads (106CFU/mL). However, the detection of lower concentrations was variable, despite the addition of pre-processing treatment to kits without such steps. Of the evaluated kits, the UMD-Universal CE IVD kit generated a higher quantity of DNA with greater nucleic acid purity and afforded the detection of the lowest microbial load in the samples. The inclusion of pre-processing steps with the kit seems to be critical for the detection of microorganism DNA directly from whole blood. In conclusion, future application of molecular techniques will require overcoming major challenges such as the detection of low levels of microorganism nucleic acids amidst the large quantity of human DNA present in samples or differences in the cellular structures of etiological agents that can also prevent high-quality DNA yields.

Does virus-bacteria coinfection increase the clinical severity of acute respiratory infection?

This retrospective cohort study investigated the presence of bacteria in respiratory secretions of patients hospitalized with acute respiratory infections and analyzed the impact of viral and bacterial coinfection on severity and the mortality rate. A total of 169 patients with acute respiratory infections were included, viruses and bacteria in respiratory samples were detected using molecular methods. Among all samples, 73.3% and 59.7% were positive for viruses and bacteria, respectively; 45% contained both virus and bacteria. Bacterial coinfection was more frequent in patients infected by community respiratory viruses than influenza A H1N1pdm (83.3% vs. 40.6%). The most frequently bacteria detected were Streptococcus pneumoniae and Haemophilus influenzae. Both species were co-detected in 54 patients and identified alone in 22 and 21 patients, respectively. Overall, there were no significant differences in the period of hospitalization, severity, or mortality rate between patients infected with respiratory viruses alone and those coinfected by viruses and bacteria. The detection of mixed respiratory pathogens is frequent in hospitalized patients with acute respiratory infections, but its impact on the clinical outcome does not appear substantial. However, it should be noted that most of the patients received broad-spectrum antibiotic therapy, which may have contributed to this favorable outcome.

Acute gastroenteritis and enteric viruses in hospitalised children in southern Brazil: aetiology, seasonality and clinical outcomes.

Viral acute gastroenteritis (AG) is a significant cause of hospitalisation in children younger than five years. Group A rotavirus (RVA) is responsible for 30% of these cases. Following the introduction of RVA immunisation in Brazil in 2006, a decreased circulation of this virus has been observed. However, AG remains an important cause of hospitalisation of paediatric patients and only limited data are available regarding the role of other enteric viruses in these cases. We conducted a prospective study of paediatric patients hospitalised for AG. Stool samples were collected to investigate human adenovirus (HAdV), RVA, norovirus (NoV) and astrovirus (AstV). NoV typing was performed by nucleotide sequencing and phylogenetic analysis. From the 225 samples tested, 60 (26%) were positive for at least one viral agent. HAdV, NoV, RVA and AstV were detected in 16%, 8%, 6% and 0% of the samples, respectively. Mixed infections were found in nine patients: HAdV/RVA (5), HAdV/NoV (3) and HAdV/NoV/RVA (1). The frequency of fever and lymphocytosis was significantly higher in virus-infected patients. Phylogenetic analysis of NoV indicated that all of these viruses belonged to genotype GII.4. The significant frequency of these pathogens in patients with AG highlights the need to routinely implement laboratory investigations.

Acute gastroenteritis and enteric viruses in hospitalised children in southern Brazil: aetiology, seasonality and clinical outcomes

Viral acute gastroenteritis (AG) is a significant cause of hospitalisation in children younger than five years. Group A rotavirus (RVA) is responsible for 30% of these cases. Following the introduction of RVA immunisation in Brazil in 2006, a decreased circulation of this virus has been observed. However, AG remains an important cause of hospitalisation of paediatric patients and only limited data are available regarding the role of other enteric viruses in these cases. We conducted a prospective study of paediatric patients hospitalised for AG. Stool samples were collected to investigate human adenovirus (HAdV), RVA, norovirus (NoV) and astrovirus (AstV). NoV typing was performed by nucleotide sequencing and phylogenetic analysis. From the 225 samples tested, 60 (26%) were positive for at least one viral agent. HAdV, NoV, RVA and AstV were detected in 16%, 8%, 6% and 0% of the samples, respectively. Mixed infections were found in nine patients: HAdV/RVA (5), HAdV/NoV (3) and HAdV/NoV/RVA (1). The frequency of fever and lymphocytosis was significantly higher in virus-infected patients. Phylogenetic analysis of NoV indicated that all of these viruses belonged to genotype GII.4. The significant frequency of these pathogens in patients with AG highlights the need to routinely implement laboratory investigations.

Caracterização molecular dos genótipos do rotavírus em pacientes pediátricos imunossuprimidos e não imunossuprimidos / Molecular characterization of rotavirus genotypes in immunosuppressed and non-immunosuppressed pediatric patients

OBJETIVO: Descrever a variabilidade genotípica do rotavírus grupo A (RVA) encontrado em pacientes pediátricos imunocompetentes e imunocomprometidos tratados no Hospital de Clínicas/Universidade Federal do Paraná (HC/UFPR), Curitiba, Paraná. MÉTODOS: Foi realizado um estudo transversal com 1.140 amostras de fezes coletadas, de abril de 2001 a dezembro de 2008, em pacientes ambulatoriais e pacientes hospitalizados com gastroenterite aguda encaminhados ao hospital. As técnicas usadas foram o método da aglutinação do látex e imunoensaio enzimático para diagnóstico de RVA. Foi realizada transcrição reversa, seguida por PCR multiplex semi-nested e sequência de nucleotídeos para caracterização do genótipo. Foram relatados dados de combinações de genótipos, clínicos, epidemiológicos, laboratoriais e sobre a presença de infecções hospitalares. RESULTADOS: Foi analisado um total de 80 amostras de fezes positivas para rotavírus. As associações mais frequentes entre os genótipos G e P foram: G4 P[8] (38,9%), G1 P[8] (30,5%), G9 P[8] (13,9%), G2 P[4] (6.9 %) e G3 P[8] 1,4%). O genótipo prevalente foi G2 P[4] depois da implementação da vacina nos anos de 2006 e 2008. Verificou-se que um total de 62,5% das crianças com idade abaixo de 12 meses estavam infectadas. Destas, 55,6% tinham grave desidratação, e 26,7% precisaram de cuidados intensivos. Encontrou-se uma frequência de 12,5% de infecções hospitalares. Não se observou correlação entre o genótipo e a gravidade da infecção nos pacientes estudados. CONCLUSÃO: As infecções por RVA podem associar-se a manifestações clínicas graves e é crucial a vigilância da variabilidade genotípica desse vírus para monitorizar a emergência de novas cepas e o impacto da imunização nesses pacientes.

OBJECTIVE: To describe the genotypic variability of group A rotavirus (RVA) found in immunosuppressed and non-immunosuppressed pediatric patients treated at the Hospital de Clínicas da Universidade Federal do Paraná (HC-UFPR), Curitiba, Paraná. METHODS: A cross-sectional study was conducted with 1,140 stool samples collected from April, 2001 to December, 2008 in outpatients and hospitalized patients with acute gastroenteritis referred to the hospital. RVA diagnosis was performed through the latex agglutination method and enzyme immunoassay. Reverse transcription followed by multiplex hemi-nested polymerase chain reaction (PCR) and nucleotide sequencing were used for genotype characterization. Genotype combinations, clinical data, epidemiological data, laboratory data, and presence of hospital-acquired infections were reported. RESULTS: A total of 80 rotavirus-positive stool samples were analyzed. The most frequent associations between genotypes G and P were: G4 P[8] (38.9%), G1 P[8] (30.5%), G9 P[8] (13.9%), G2 P[4] (6.9%), and G3 P[8] (1.4%). G2 P[4] was the most prevalent genotype after the vaccine implementation in the years 2006 and 2008. A total of 62.5% of children aged less than 12 months were found to be infected. Of these, 55.6% had severe dehydration and 26.7% needed intensive care. A frequency of 12.5% of nosocomial infections was found. No correlation was observed between genotype and severity of infection in the study patients. CONCLUSION: RVA infections can be associated with severe clinical manifestations, and the surveillance of genotypic variability of this virus is crucial to monitor the emergence of new strains and the impact of the immunization in these patients.

Molecular characterization of rotavirus genotypes in immunosuppressed and non-immunosuppressed pediatric patients.

OBJECTIVE: To describe the genotypic variability of group A rotavirus (RVA) found in immunosuppressed and non-immunosuppressed pediatric patients treated at the Hospital de Clínicas da Universidade Federal do Paraná (HC-UFPR), Curitiba, Paraná. METHODS: A cross-sectional study was conducted with 1,140 stool samples collected from April, 2001 to December, 2008 in outpatients and hospitalized patients with acute gastroenteritis referred to the hospital. RVA diagnosis was performed through the latex agglutination method and enzyme immunoassay. Reverse transcription followed by multiplex hemi-nested polymerase chain reaction (PCR) and nucleotide sequencing were used for genotype characterization. Genotype combinations, clinical, epidemiological, laboratory data, and presence of hospital-acquired infections were reported. RESULTS: A total of 80 rotavirus-positive stool samples were analyzed. The most frequent associations between genotypes G and P were: G4 P[8] (38.9%), G1 P[8] (30.5%), G9 P[8] (13.9%), G2 P[4] (6.9%), and G3 P[8] (1.4%). G2 P[4] was the most prevalent genotype after the vaccine implementation in the years 2006 and 2008. A total of 62,5% of infected children were aged less than 12 months. Of these, 55.6% had severe dehydration and 26.7% needed intensive care. A frequency of 12.5% of nosocomial infections was found. No correlation was observed between genotype and severity of infection in the study patients. CONCLUSION: RVA infections can be associated with severe clinical manifestations, and the surveillance of genotypic variability of this virus is crucial to monitor the emergence of new strains and the impact of the immunization in these patients.

Impact of human metapneumovirus infection on in and outpatients for the years 2006-2008 in Southern Brazil

The human metapneumovirus (hMPV), member of the Paramyxoviridae family, has been reported as an important agent involved with acute respiratory infections (ARIs). The aim of this study is to identify hMPV as the etiological agent of ARIs on in and outpatients in the city of Curitiba, Southern Brazil, and describe clinical data of hMPV subtyping. A retrospective study was performed in 1,572 respiratory samples over a period of three years. hMPV was detected by reverse transcription-polymerase chain reaction and subtyping was performed by nucleotide sequencing. hMPV was present in 61 (3.9 percent) samples and subtypes A1, A2a, B1 and B2 were detected. The incidence of hMPV was higher in outpatients (5.9 percent), whose mean age was 19.7 years (range 6 months-75 years old), than in inpatients (3 percent), whose mean age was 7.6 months (range 1 month-26 years old). The outpatients had upper respiratory tract infections with flu-like symptoms and all hospitalized children had lower respiratory tract infections. A pediatric patient died from complications associated with hMPV A2a infection. hMPV has been reported as a respiratory pathogen in all age groups. No correlation was observed between viral subtype and disease severity in the samples of this study.

Adenovirus respiratory infection: significant increase in diagnosis using PCR comparing with antigen detection and culture methods / Infecção respiratória aguda por adenovirus: comparação dos métodos de PCR e imunofluorescência indireta para o seu diagnóstico e dados clínicos dos pacientes infectados

Adenovirus (AdV) respiratory infections are usually described as being associated with high mortality rates. Laboratory diagnosis is essential for the establishment of the appropriate therapy, and for guiding the implementation of preventive measures in order to prevent the spread of the infection. Aiming to analyze the sensitivity and specificity of the laboratorial diagnosis methods available, we compared antigen detection by indirect immunofluorescence assay (IF), and a specific nested polymerase chain reaction (PCR), to detect AdV in respiratory samples collected from patients admitted to hospital with acute respiratory disease. Positive samples were inoculated into a cell culture to confirm the results. We analyzed 381 samples from the nasopharyngeal aspirates collected during the year 2008; of these, 2.6 percent tested were positive for adenovirus through IF and 10 percent through PCR; positive isolation was obtained in 40 percent and 26 percent of these cases, respectively. Most infected patients were children under six months of age, and despite of the fact that a significant number of patients required intensive care, the mortality rate was low (5 percent). In conclusion, molecular methods were found to be useful for rapid diagnosis of adenovirus infections with higher sensitivity than antigen detection; their introduction permitted a significant increase in diagnoses of adenovirus infections.

Infecções respiratórias por Adenovírus (ADV) são geralmente descritas associadas com alta mortalidade. O diagnóstico laboratorial é essencial para o estabelecimento da terapêutica adequada e para orientar a implantação de medidas preventivas evitando a propagação da infecção. Com o objetivo de analisar a sensibilidade e a especificidade dos métodos de avaliação de diagnóstico laboratorial, foi comparada a detecção de antígeno por imunofluorescência indireta (IF) com a reação em cadeia da polimerase específica (PCR) para detectar AdV em amostras respiratórias coletadas de pacientes internados com doença respiratória aguda. As amostras com resultados positivos foram inoculadas em cultura celular. Foram analisadas 381 amostras da secreção nasofaríngea coletadas durante o ano de 2008, das quais 2,6 por cento foram positivas pela IF e 10 por cento pela PCR, isolamento positivo foi obtido em 40 por cento e 26 por cento dos casos positivos pelos testes anteriores, respectivamente. A maioria dos pacientes infectados eram crianças com menos de seis meses de idade, e apesar do fato de que um número significativo de pacientes necessitou de cuidados intensivos, a taxa de mortalidade foi baixa (5 por cento). Em conclusão, os métodos moleculares são úteis para o diagnóstico rápido de infecções por adenovírus com maior sensibilidade do que a detecção do antígeno, a sua introdução na rotina permitiu um aumento significativo no diagnóstico de infecções por adenovírus.

Impact of human metapneumovirus infection on in and outpatients for the years 2006-2008 in Southern Brazil.

The human metapneumovirus (hMPV), member of the Paramyxoviridae family, has been reported as an important agent involved with acute respiratory infections (ARIs). The aim of this study is to identify hMPV as the etiological agent of ARIs on in and outpatients in the city of Curitiba, Southern Brazil, and describe clinical data of hMPV subtyping. A retrospective study was performed in 1,572 respiratory samples over a period of three years. hMPV was detected by reverse transcription-polymerase chain reaction and subtyping was performed by nucleotide sequencing. hMPV was present in 61 (3.9%) samples and subtypes A1, A2a, B1 and B2 were detected. The incidence of hMPV was higher in outpatients (5.9%), whose mean age was 19.7 years (range 6 months-75 years old), than in inpatients (3%), whose mean age was 7.6 months (range 1 month-26 years old). The outpatients had upper respiratory tract infections with flu-like symptoms and all hospitalized children had lower respiratory tract infections. A pediatric patient died from complications associated with hMPV A2a infection. hMPV has been reported as a respiratory pathogen in all age groups. No correlation was observed between viral subtype and disease severity in the samples of this study.

Adenovirus respiratory infection: significant increase in diagnosis using PCR comparing with antigen detection and culture methods.

Adenovirus (AdV) respiratory infections are usually described as being associated with high mortality rates. Laboratory diagnosis is essential for the establishment of the appropriate therapy, and for guiding the implementation of preventive measures in order to prevent the spread of the infection. Aiming to analyze the sensitivity and specificity of the laboratorial diagnosis methods available, we compared antigen detection by indirect immunofluorescence assay (IF), and a specific nested polymerase chain reaction (PCR), to detect AdV in respiratory samples collected from patients admitted to hospital with acute respiratory disease. Positive samples were inoculated into a cell culture to confirm the results. We analyzed 381 samples from the nasopharyngeal aspirates collected during the year 2008; of these, 2.6% tested were positive for adenovirus through IF and 10% through PCR; positive isolation was obtained in 40% and 26% of these cases, respectively. Most infected patients were children under six months of age, and despite of the fact that a significant number of patients required intensive care, the mortality rate was low (5%). In conclusion, molecular methods were found to be useful for rapid diagnosis of adenovirus infections with higher sensitivity than antigen detection; their introduction permitted a significant increase in diagnoses of adenovirus infections.

The epidemiology and antigenic characterization of influenza viruses isolated in Curitiba, South Brazil.

Several studies conducted all over the world have reported that the influenza virus is associated with great morbidity and mortality rates. In this study, we analyzed the incidence of the influenza virus between 2000 and 2003 in Curitiba. We studied 1621 samples obtained from outpatients and hospitalized patients of both sexes and all ages. The study was conducted at the local primary care health units (outpatients) and at the tertiary care unit (hospitalized) of the General Hospital of the Federal University in the state of Paraná, Brazil. Nasopharyngeal aspirates and, eventually, bronchoalveolar lavage were assayed for the presence of viral antigens, either by indirect immunofluorescence or cell culture. Of the samples studied, 135 (8.3%) were positive for influenza virus, and of those, 103 (76.3%) were positive for type A and 32 (23.7%) for type B. Additionally, positive samples were analyzed by reverse transcription followed by polymerase chain reaction and subtypes H1 and H3 were identified from this group. A high incidence of positive samples was observed mainly in the months with lower temperatures. Furthermore, outpatients showed a higher incidence of influenza viruses than hospitalized patients.

The epidemiology and antigenic characterization of influenza viruses isolated in Curitiba, South Brazil

Several studies conducted all over the world have reported that the influenza virus is associated with great morbidity and mortality rates. In this study, we analyzed the incidence of the influenza virus between 2000 and 2003 in Curitiba. We studied 1621 samples obtained from outpatients and hospitalized patients of both sexes and all ages. The study was conducted at the local primary care health units (outpatients) and at the tertiary care unit (hospitalized) of the General Hospital of the Federal University in the state of Paraná, Brazil. Nasopharyngeal aspirates and, eventually, bronchoalveolar lavage were assayed for the presence of viral antigens, either by indirect immunofluorescence or cell culture. Of the samples studied, 135 (8.3 percent) were positive for influenza virus, and of those, 103 (76.3 percent) were positive for type A and 32 (23.7 percent) for type B. Additionally, positive samples were analyzed by reverse transcription followed by polymerase chain reaction and subtypes H1 and H3 were identified from this group. A high incidence of positive samples was observed mainly in the months with lower temperatures. Furthermore, outpatients showed a higher incidence of influenza viruses than hospitalized patients.

Relação entre a carga viral de HPV oncogênico determinada pelo método de captura híbrida e o diagnóstico citológico de lesões de alto grau / Association between high-risk human papillomavirus DNA load detected by hybrid capture II and high-grade precursor lesions of cervical cancer in women

INTRODUÇÃO: O papilomavírus humano (HPV) é o principal fator de risco para as neoplasias intra-epiteliais cervicais (NIC) e o câncer cervical. OBJETIVO: O objetivo deste estudo é avaliar se há associação entre a carga viral de HPV oncogênico (alto risco), determinada por meio do teste molecular captura híbrida II (CH II), e o diagnóstico de lesões de alto grau (NIC II/III). MATERIAL E MÉTODOS: Foram analisadas 982 amostras cervicovaginais de exames ginecológicos de rotina, obtidas pelos métodos Papanicolaou convencional e/ou citologia em base líquida (DNA-Citoliq-Digene). Os resultados foram confirmados utilizando-se o método de captura híbrida (CH [Digene]) para detecção de DNA/HPV de alto grau. Os resultados com valor > 1 pg/ml foram considerados positivos, e esses foram divididos em dois grupos: 1. carga viral < 100 pg/ml, e 2. carga viral > 100 pg/ml. RESULTADOS: Dos 210 (21,4 por cento) casos diagnosticados como NIC I, 152 (72,4 por cento) foram positivos para HPV de alto risco por CH II. Desses, 101 (66,4 por cento) apresentaram carga viral > 100 pg/ml. O diagnóstico de NIC II ou III foi confirmado por CH II de alto risco em 86 (43,6 por cento) casos, contudo, entre esses, em 53 (61,6 por cento) a carga viral detectada foi > 100 pg/ml. DISCUSSÃO E CONCLUSÃO: Nossos resultados demonstram que há uma clara associação entre o valor da carga viral determinada pelo método CH II (versão 1) e o grau das lesões precursoras de câncer. Pacientes com carga viral superior a 100 'g/ml devem ser monitoradas periodicamente.

INTRODUCTION: Infection with oncogenic human papilloma virus (HPV) has been established as the main etiologic agent for cervical cancer and of cervical intraepithelial neoplasia (CIN). OBJECTIVE: To determine the association between viral loads of the high risk HPV using the hybrid capture II (HC II) system and CIN lesion stage. MATERIAL AND METHOD: A total of 982 women with diagnosis of negative or of CIN I-III with Pap or liquid-based cytology (DNA-Citoliq-Digene) were included. HC II testing was used to detected HPV DNA. Viral load was measured expressed as relative lights unit (RLU) ratio. RLU ratios were categorized for analysis into three groups: negative (< 1 pg/ml); positive with low viral load (< 100 pg/ml); positive with high viral load (> 100 pg/ml). RESULTS: A total of 152 (72.4 percent) of 210 (21.4 percent) patients with CIN II, 53 (61.6 percent) of 86 (43.6 percent) with CIN II-III were positive for HPV DNA, the higher the viral load (> 100 pg/ml) were observed in 101 (66.4 percent) and 53 (61.6 percent) patients with CIN I or CIN II-III, respectively. DISCUSSION AND CONCLUSIONS: Our results suggest a clear association between viral load HPV DNA determined by HC II (version 1) assay and CIN stage. We concluded that the high viral load has a clinical importance for the management of patients with CIN.

Viral respiratory infection in Curitiba, Southern Brazil.

OBJECTIVE: Viral infection is the major cause of respiratory tract pathology affecting individuals of all ages, especially children and immunocompromised patients. There is a paucity of data on etiological and epidemiological infection caused by virus, in Southern Brazil. The aim of this study was to evaluate the seasonality, and the incidence of the viruses involved in respiratory infections in Curitiba, South Brazil. METHODS: Two hundred seventy-three nasopharyngeal aspirates (NPA) from primary care health units and 1348 NPA or bronchoalveolar lavages from a tertiary-care teaching hospital were studied. Viruses were identified by indirect immunofluorescence and cellular culture. RESULTS: Thirty percent of tested samples scored positive for the presence of virions. The percentages of infection for each virus from outpatients and hospitalized, respectively, were 3.3, 19.3 for respiratory syncytial virus; 13.9, 3.5 for influenza A, 3.0, 1.7 for influenza B; 3.3, 5.6 for parainfluenza virus and 2.2, 0.8 for adenovirus. A higher incidence of positive results was found during the winter season, thus showing a pattern of seasonality. CONCLUSION: Viral agents are one of the main etiologies of respiratory tract pathology in the population studied.