ABSTRACT Purpose: This study aimed to evaluate the pre-sence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in the ocular surface of individuals clinically suspected of coronavirus disease 2019 (COVID-19) and determine the accuracy of different approaches of molecular testing on the ocular surface based on the nasopharyngeal positivity status for COVID-19. Methods: A total of 152 individuals with suspected COVID-19 symptoms who simultaneously underwent nasopharyngeal and two different tear film collection techniques for quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) were included. Tears were collected and randomized: one eye had the filter strip for the Schirmer test and the contralateral eye had conjunctival swab/cytology in the inferior fornix. All patients underwent slit lamp biomicroscopy. The accuracy of various ocular surface collection techniques used for the detection of SARS-CoV-2 RNA was determined. Results: Of the 152 patients enrolled in the study, 86 (56.6%) had COVID-19 confirmed by nasopharyngeal PCR. Both tear film collection techniques detected viral particles: the Schirmer test was positive in 16.3% (14/86) and the conjunctival swab/cytology in 17.4% (15/86), with no statistically significant differences. No positive ocular tests were found among those with negative nasopharyngeal PCR tests. The overall agreement of the ocular tests was 92.7%, and in combination, the sensitivity would increase to 23.2%. The mean cycle threshold values in the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests were 18.2 ± 5.3, 35.6 ± 1.4, and 36.4 ± 3.9, respectively. Compared with the nasopharyngeal test, the Schirmer (p=0.001) and conjunctival swab/cytology (p<0.001) tests had significantly different Ct values. Conclusion: The Schirmer (16.3%) and conjunctival swab (17.4%) tests were comparably capable of detecting SARS-CoV-2 RNA in the ocular surface by RT-PCR accurately based on nasopharyngeal status and demonstrated indistinct sensitivity and specificity. Simultaneous specimen sampling and processing from the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests demonstrated significantly lower viral load in both ocular surface approaches than in the nasopharyngeal test. Ocular manifestations detected by slit lamp biomicroscopy were not associated with ocular RT-PCR positivity.

RESUMO Objetivo: Avaliar a presença de RNA de coronavírus 2 causador de síndrome respiratória aguda grave (SARS-CoV-2) na superfície ocular de indivíduos clinicamente suspeitos com COVID-19 e determinar a precisão de diferentes abordagens de testes moleculares na superfície ocular com base no status de positividade do RT-qPCR de nasofaringe para COVID-19. Métodos: 152 indivíduos com sintomas suspeitos para a COVID-19 foram submetidos a coleta de reação em cadeia da polimerase de nasofaringe simultaneamente a duas técnicas diferentes de coleta de filme lacrimal para RT-qPCR: aleatoriamente, um olho com a tira filtro do teste de Schirmer e, o olho contralateral, com citologia (swab) conjuntival no fórnice inferior. Todos os indivíduos foram submetidos à biomicroscopia com lâmpada de fenda. Resultados: Dos 152 pacientes, 86 (56,6%) tiveram a COVID-19 confirmada por PCR de nasofaringe. Ambas as técnicas de coleta detectaram partículas virais: o teste de Schirmer foi positivo em 16,3% (14/86) e a citologia conjuntival em 17,4% (15/86), sem diferenças estatisticamente significativas. Não houve testes oculares positivos entre aqueles com reação em cadeia da polimerase de nasofaringe negativo. A concordância geral dos testes oculares foi de 92,7% e, em combinação, a sensibilidade aumentaria para 23,2%. Os valores médios do limiar de ciclo nos testes de nasofaringe, Schirmer e citologia conjuntival foram 18,2 ± 5,3, 35,6 ± 1,4 e 36,4 ± 3,9, respectivamente. Conclusão: Os testes de Schirmer (16,3%) e swab conjuntival (17,4%) foram igualmente capazes de detectar RNA de SARS-CoV-2 na superfície ocular por RT-PCR e demonstraram sensibilidade e especificidade indistintas. A coleta simultânea de amostras ao processamento dos testes de RT-PCR de nasofaringe, Schirmer e citologia (swab) conjuntival demonstraram carga viral significativamente menor em ambas as abordagens da superfície ocular em comparação com o teste de nasofaringe. As manifestações oculares detectadas pela biomicroscopia com lâmpada de fenda não foram claramente associadas à positividade do RT-PCR ocular.

RESUMO Objetivo: Avaliar a presença de RNA de coronavírus 2 causador de síndrome respiratória aguda grave (SARS-CoV-2) na superfície ocular de indivíduos clinicamente suspeitos com COVID-19 e determinar a precisão de diferentes abordagens de testes moleculares na superfície ocular com base no status de positividade do RT-qPCR de nasofaringe para COVID-19. Métodos: 152 indivíduos com sintomas suspeitos para a COVID-19 foram submetidos a coleta de reação em cadeia da polimerase de nasofaringe simultaneamente a duas técnicas diferentes de coleta de filme lacrimal para RT-qPCR: aleatoriamente, um olho com a tira filtro do teste de Schirmer e, o olho contralateral, com citologia (swab) conjuntival no fórnice inferior. Todos os indivíduos foram submetidos à biomicroscopia com lâmpada de fenda. Resultados: Dos 152 pacientes, 86 (56,6%) tiveram a COVID-19 confirmada por PCR de nasofaringe. Ambas as técnicas de coleta detectaram partículas virais: o teste de Schirmer foi positivo em 16,3% (14/86) e a citologia conjuntival em 17,4% (15/86), sem diferenças estatisticamente significativas. Não houve testes oculares positivos entre aqueles com reação em cadeia da polimerase de nasofaringe negativo. A concordância geral dos testes oculares foi de 92,7% e, em combinação, a sensibilidade aumentaria para 23,2%. Os valores médios do limiar de ciclo nos testes de nasofaringe, Schirmer e citologia conjuntival foram 18,2 ± 5,3, 35,6 ± 1,4 e 36,4 ± 3,9, respectivamente. Conclusão: Os testes de Schirmer (16,3%) e swab conjuntival (17,4%) foram igualmente capazes de detectar RNA de SARS-CoV-2 na superfície ocular por RT-PCR e demonstraram sensibilidade e especificidade indistintas. A coleta simultânea de amostras ao processamento dos testes de RT-PCR de nasofaringe, Schirmer e citologia (swab) conjuntival demonstraram carga viral significativamente menor em ambas as abordagens da superfície ocular em comparação com o teste de nasofaringe. As manifestações oculares detectadas pela biomicroscopia com lâmpada de fenda não foram claramente associadas à positividade do RT-PCR ocular.

ABSTRACT Purpose: This study aimed to evaluate the pre-sence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in the ocular surface of individuals clinically suspected of coronavirus disease 2019 (COVID-19) and determine the accuracy of different approaches of molecular testing on the ocular surface based on the nasopharyngeal positivity status for COVID-19. Methods: A total of 152 individuals with suspected COVID-19 symptoms who simultaneously underwent nasopharyngeal and two different tear film collection techniques for quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) were included. Tears were collected and randomized: one eye had the filter strip for the Schirmer test and the contralateral eye had conjunctival swab/cytology in the inferior fornix. All patients underwent slit lamp biomicroscopy. The accuracy of various ocular surface collection techniques used for the detection of SARS-CoV-2 RNA was determined. Results: Of the 152 patients enrolled in the study, 86 (56.6%) had COVID-19 confirmed by nasopharyngeal PCR. Both tear film collection techniques detected viral particles: the Schirmer test was positive in 16.3% (14/86) and the conjunctival swab/cytology in 17.4% (15/86), with no statistically significant differences. No positive ocular tests were found among those with negative nasopharyngeal PCR tests. The overall agreement of the ocular tests was 92.7%, and in combination, the sensitivity would increase to 23.2%. The mean cycle threshold values in the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests were 18.2 ± 5.3, 35.6 ± 1.4, and 36.4 ± 3.9, respectively. Compared with the nasopharyngeal test, the Schirmer (p=0.001) and conjunctival swab/cytology (p<0.001) tests had significantly different Ct values. Conclusion: The Schirmer (16.3%) and conjunctival swab (17.4%) tests were comparably capable of detecting SARS-CoV-2 RNA in the ocular surface by RT-PCR accurately based on nasopharyngeal status and demonstrated indistinct sensitivity and specificity. Simultaneous specimen sampling and processing from the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests demonstrated significantly lower viral load in both ocular surface approaches than in the nasopharyngeal test. Ocular manifestations detected by slit lamp biomicroscopy were not associated with ocular RT-PCR positivity.

Abstract Background The neurological manifestations in COVID-19 adversely impact acute illness and post-disease quality of life. Limited data exist regarding the association of neurological symptoms and comorbid individuals. Objective To assess neurological symptoms in hospitalized patients with acute COVID-19 and multicomorbidities. Methods Between June 2020 and July 2020, inpatients aged 18 or older, with laboratory-confirmed COVID-19, admitted to the Hospital São Paulo (Federal University of São Paulo), a tertiary referral center for high complexity cases, were questioned about neurological symptoms. The Composite Autonomic Symptom Score 31 (COMPASS-31) questionnaire was used. The data were analyzed as a whole and whether subjective olfactory dysfunction was present or not. Results The mean age of the sample was 55 ± 15.12 years, and 58 patients were male. The neurological symptoms were mostly xerostomia (71%), ageusia/hypogeusia (50%), orthostatic intolerance (49%), anosmia/hyposmia (44%), myalgia (31%), dizziness (24%), xerophthalmia (20%), impaired consciousness (18%), and headache (16%). Furthermore, 91% of the patients had a premorbidity. The 44 patients with subjective olfactory dysfunction were more likely to have hypertension, diabetes, weakness, shortness of breath, ageusia/hypogeusia, dizziness, orthostatic intolerance, and xerophthalmia. The COMPASS-31 score was higher than that of previously published controls (14.85 ± 12.06 vs. 8.9 ± 8.7). The frequency of orthostatic intolerance was 49% in sample and 63.6% in those with subjective olfactory dysfunction (2.9-fold higher risk compared to those without). Conclusion A total of 80% of inpatients with multimorbidity and acute COVID-19 had neurological symptoms. Chemical sense and autonomic symptoms stood out. Orthostatic intolerance occurred in around two-thirds of the patients with anosmia/hyposmia. Hypertension and diabetes were common, mainly in those with anosmia/hyposmia.

Resumo Antecedentes As manifestações neurológicas na COVID-19 impactam adversamente na enfermidade aguda e na qualidade de vida após a doença. Dados limitados existem em relação a associação de sintomas neurológicos e indivíduos com comorbidades. Objetivo Avaliar os sintomas neurológicos em pacientes de hospitalizados com COVID-19 aguda e múltiplas comorbidades. Métodos Entre junho e julho de 2020, pacientes de hospitais com idade 18 anos ou acima e COVID-19 laboratorialmente confirmada, admitidos no Hospital São Paulo (Universidade Federal de São Paulo), um centro de referência terciário para casos de alta complexidade, foram perguntados sobre sintomas neurológicos. O questionário Pontuação composta de sintoma autonômico (COMPASS-31) foi usado. Os dados foram analisados no geral e se a disfunção olfatória subjetiva estava presente ou não. Resultados A média de idade da amostra foi 55 ± 15.12 anos. 58 pacientes eram homens. Os sintomas neurológicos foram principalmente xerostomia (71%), ageusia/hipogeusia (50%), intolerância ortostática (49%), anosmia/hiposmia (44%), mialgia (31%), tontura (24%), xeroftalmia (20%), comprometimento na consciência (18%) e cefaleia (16%). Além disso, 91 % dos pacientes tinham uma pré-morbidade. Os 44 pacientes com disfunção olfatória tinham maior chance de ter hipertensão, diabetes, fraqueza, falta de ar, ageusia/hipogeusia, tontura, intolerância ortostática e xeroftalmia. A pontuação do COMPASS-31 foi maior do que a de controles previamente publicados (14,85 ± 12,06 vs. 8,9 ± 8,7). A frequência de intolerância ortostática foi 49% na amostra e 63,6% naqueles com disfunção olfatória subjetiva (risco 2.9 vezes maior comparado com os sem). Conclusão Um total de 80% dos pacientes hospitalizados com múltiplas morbidades e COVID-19 aguda tinham sintomas neurológicos. Os sintomas do sentido químico e autonômicos se destacaram. A intolerância ortostática ocorreu em cerca de dois terços dos pacientes com anosmia/hiposmia. A hipertensão e o diabetes foram comuns, principalmente naqueles com anosmia/hiposmia.

OBJETIVO: Os adenovírus desempenham um papel importante na etiologia da infecção aguda grave do trato respiratório inferior, especialmente entre crianças. O objetivo do estudo foi avaliar a detecção do adenovírus humano (HAdV) por diferentes métodos (imunofluorescência direta DFA e reação em cadeia da polimerase nested nested PCR) em amostras coletadas de diferentes populações de pacientes pediátricos. MÉTODOS: O material foi coletado de crianças portadoras de doença cardíaca congênita (DCC 123 aspirados nasais coletados em 2005, 2007 e 2008) e de crianças da comunidade (CC 165 aspirados nasais coletados em 2008). As crianças eram consideradas elegíveis se apresentassem infecção respiratória aguda (IRA) de provável etiologia viral, com até sete dias de início dos sintomas. Todas as amostras coletadas no estudo foram avaliadas por meio de DFA e nested PCR. RESULTADOS: De 209 amostras incluídas, 43 (14,8%) foram positivas em pelo menos um dos testes feitos: 17/165 (10,3%) das crianças da comunidade e 26/125 (20,8%) das crianças cardiopatas. As taxas de detecção por nested PCR foram 15/165 (9,1%) em crianças da comunidade e 24/125 (19,2%) em crianças cardiopatas. O método molecular mostrou maiores taxas de detecção quando comparado com a DFA (p<0,001). A análise univariada mostrou que as crianças portadoras de cardiopatia congênita apresentaram chance significativamente maior de adquirir HAdV (odds ratio 2,3; IC 95%: 1,18-4,43). CONCLUSÕES: Baseado nos resultados obtidos na presente avaliação, recomenda-se a vigilância de rotina em pacientes de risco (DCC) por métodos moleculares, que melhora o fluxo diagnóstico e a eficiência da detecção.

OBJECTIVE: Adenoviruses play an important role in the etiology of severe acute lower respiratory infection, especially in young children. The aim of the present study was to evaluate the Human Adenovirus (HAdV) detection by different methods (Direct Fluorescence Assay DFA and Nested Polymerase Chain Reaction nested PCR), among samples collected from different groups of pediatric patients. METHODS: Collection of samples was made in children with congenital heart disease (CHD 123 nasal aspirates collected in the years of 2005, 2007 and 2008) and in community children (CC 165 nasal aspirates collected in 2008). Children were eligible if they presented acute respiratory infection (ARI) of probable viral etiology, within up to 7 days of symptoms' onset. All studied samples were evaluated by DFA and nested PCR assay. RESULTS: Of the 290 samples included during the study period, 43 (14.8%) were positive on at least one test: 17/165 (10.3%) of the CC and 26/125 (20.8%) of the CHD children. The nested PCR detection rates in the community children were 15/165 (9.1%), and for children with CHD, 24/125 (19.2%). Molecular method showed higher detection rates when compared to the DFA test (p<0.001). Univariate analysis showed that children with congenital heart disease presented a significantly higher chance for acquiring the HAdV (Odds Ratio 2.3; 95% CI: 1.18-4.43). CONCLUSIONS: Based on data obtained in the present evaluation, we suggest that a routine surveillance should be performed in high risk patients by molecular methods, thus improving diagnostic flow and efficiency.

OBJECTIVES: The pandemic of 2009 H1N1 influenza A emerged in February 2009, with high morbidity and mortality, and rapidly spread globally. São Paulo was among the most affected areas in Brazil. This study compares the clinical and epidemiological characteristics of influenza-like illness between outpatients and hospitalized patients and evaluates the impact of oseltamivir therapy on the outcome of 2009 H1N1 influenza A patients. METHODS: This is a case series study comparing the clinical and epidemiological characteristics of influenza-like illness between outpatients attended at Hospital São Paulo in August 2009 (the peak of the first pandemic wave) and those patients hospitalized between May and September 2009 (the entire first pandemic wave). RESULTS: The 1651 patients evaluated were predominantly female (927×686, p<0.001) and aged 31.71±16.42 years, with 148 reporting chronic pulmonary disease. Dyspnea was presented by 381 (23.4%) patients and was more frequent among those aged 30 years or more (p<0.001). Hospitalization occurred at 3.73±2.85 days, and antiviral treatment started 2.27±2.97 days after the onset of first symptoms. A delay of more than 5 days in starting oseltamivir therapy was independently associated with hospitalization (p<0.001), a stay in the ICU (p<0.001) and a higher risk of dying (OR = 28.1, 95% CI 2.81-280.2, p = 0.007). CONCLUSION: The 2009 pandemic of H1N1 influenza A affected young adults, presented a significant disease burden and produced severe cases with a significant fatality rate. However, promptly starting specific therapy improved the outcome.

lIn 2009, the influenza A (H1N1) virus spread rapidly around the world, causing the first pandemic of the 21st Century. In 2010, there was a vaccination campaign against this new virus subtype to reduce the morbidity and mortality of the disease in some countries, including Brazil. Herein, we describe the clinical and epidemiological characteristics of patients under 19 years of age who were hospitalized with confirmed influenza A (H1N1) infection in 2009 and 2010. We retrospectively reviewed files from the pediatric patients who were admitted to a university hospital with real-time polymerase chain reaction (RT-PCR) confirmed influenza A (H1N1) infection in 2009 and 2010. There were 37 hospitalized patients with influenza A (H1N1) in 2009 and 2 in 2010. In 2009, many of the hospitalized children had an underlying chronic disease and a lower median age than those not hospitalized. Of the hospitalized patients, 78% had a chronic disease, primarily pneumopathy (48%). The main signs and symptoms of influenza were fever (97%), cough (76%), and dyspnea (59%). Complications occurred in 81% of the patients. The median length of hospitalization was five days; 27% of the patients required intensive care, and two died. In 2010, two patients were hospitalized with influenza A (H1N1): one infant with adenovirus co-infection who had received one previous H1N1 vaccine dose and presented with respiratory sequelae and a 2-month-old infant who had a hospital-acquired infection. An impressive reduction in hospital admissions was observed in 2010 when the vaccination campaign took place in Brazil.