Severe acute respiratory syndrome coronavirus 2 infection among healthcare workers in a tertiary public hospital in Curitiba, Brazil

ABSTRACT BACKGROUND: We aimed to describe the clinical characteristics of coronavirus disease 2019 (COVID-19) among healthcare workers (HCWs) in Curitiba, Brazil. METHODS: Upper respiratory samples from 1077 HCWs were tested for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using reverse transcription polymerase chain reaction from June 16, 2020 to December 9, 2020. RESULTS: Overall, 32.7% of HCWs were infected. The positivity rates in symptomatic and asymptomatic HCWs were 39.2% and 15.9%, respectively. Hospital departments categorized as high-risk for exposure had the highest number of infected HCWs. CONCLUSIONS: Early diagnosis and isolation of infected HCWs remain key in controlling SARS-CoV-2 transmission because HCWs in close contact with COVID-19 patients are more likely to be infected than those who are not.

Higher positive identification of malignant CSF cells using the cytocentrifuge than the Suta chamber / A identificação de células neoplásicas no LCR foi maior com o uso da citocentrifuga do que com câmara de Suta

ABSTRACT Objective To define how to best handle cerebrospinal fluid (CSF) specimens to obtain the highest positivity rate for the diagnosis of malignancy, comparing two different methods of cell concentration, sedimentation and cytocentrifugation. Methods A retrospective analysis of 411 CSF reports. Results This is a descriptive comparative study. The positive identification of malignant CSF cells was higher using the centrifuge than that using the Suta chamber (27.8% vs. 19.0%, respectively; p = 0.038). Centrifuge positively identified higher numbers of malignant cells in samples with a normal concentration of white blood cells (WBCs) (< 5 cells/mm3) and with more than 200 cells/mm3, although this was not statistically significant. There was no lymphocyte loss using either method. Conclusions Cytocentrifugation positively identified a greater number of malignant cells in the CSF than cytosedimentation with the Suta chamber. However, there was no difference between the methods when the WBC counts were within the normal range.

RESUMO Objetivo Definir qual a melhor forma de concentrar amostras de LCR para obter maior porcentagem de positividade para o diagnóstico de infiltração neoplásica. comparando dois métodos diferentes de concentração de células, sedimentação e citocentrifugação. Métodos Análise retrospectiva de 411 laudos de LCR. Resultados Estudo comparativo descritivo. A identificação de células neoplásicas no LCR foi mais elevada quando usada a citocentrífuga do que a câmara de Suta (28% vs 19,0%, respectivamente; p = 0,038). Centrifugação identificou maior número de células neoplásicas em amostras com concentração de células < 5 células/mm3 e superior a 200 células/mm3, embora não significativo. Não houve perda de linfócitos usando qualquer um dos métodos. Conclusões A citocentrifugação identificou um número maior de células malignas no LCR do que a sedimentação com a câmara de Suta. No entanto, não houve diferença entre os métodos quando as contagens de leucócitos estavam dentro do intervalo normal.

Cerebrospinal fluid can be used for HIV genotyping when it fails in blood / O líquido cefalorraquidiano pode ser usado para a genotipagem do HIV quando esta não é possível no plasma

Blood plasma specimens are the clinical standard for HIV-1 pol gene genotyping from viral populations; however, it is not always successful, often from low viral loads or the presence of polymerase chain reaction (PCR) inhibitors. Objective To describe the successful of HIV-1 genotyping in two samples of cerebrospinal fluid (CSF), after genotype procedures failed from blood. Method Two HIV-infected patients enrolled in a neurocognitive research study were evaluated when standard HIV-1 genotyping failed from blood plasma samples. Genotyping was performed using the commercial system TRUGENE® HIV-1 Genotyping Kit and the OpenGene® DNA Sequencing System (Siemens Healthcare Diagnostics, Tarrytown, NY, USA). Results CSF genotyping was performed via the same commercial platform and was successful in both cases. Conclusion This report demonstrates that CSF could be used as an alternate clinical specimen for HIV-1 genotyping when it fails from blood. .

O plasma é a amostra clínica padrão utilizada para a genotipagem da região pol do HIV-1; entretanto, a genotipagem pode nem sempre ser bem sucedida, geralmente devido a baixas cargas virais ou à presença de inibidores da reação em cadeia da polimerase (PCR). Objetivo: Descrever o sucesso da genotipagem do HIV-1 em duas amostras de líquido cefalorraquidiano (LCR) após a falha do mesmo método em amostras de plasma dos mesmos pacientes. Método: Dois pacientes HIV+ envolvidos em um estudo neurocognitivo foram avaliados após a falha da genotipagem do HIV-1 no plasma. A genotipagem foi realizada com o sistema comercial TRUGENE® HIV-1 Genotyping e o OpenGene® DNA Sequencing (Siemens Healthcare Diagnostics, Tarrytown, NY, USA). Resultados: A genotipagem no LCR foi realizada pelo mesmo método utilizado no plasma, sendo bem sucedida para ambos os pacientes. Conclusão: Este artigo demonstra que o LCR pode ser usado como uma amostra clínica alternativa para a genotipagem do HIV-1 quando esta falha no plasma. .

Genotypical diversity of HIV clades and central nervous system impairment / Diversidade genotípica dos subtipos de HIV e comprometimento do sistema nervoso central

The central nervous system (CNS) and the immune system are considered major target organs for HIV infection. The neurological manifestations directly related to HIV are acute viral meningitis, chronic meningitis, HIV associated dementia, vacuolar myelopathy and involvement of the peripheral nervous system. Changes in diagnosis and clinical management have changed the aspect of HIV infection so that it is no longer a fatal disease, and has become a chronic disease requiring sustained medical management. After HAART the incidence of most opportunistic infections, including those affecting the CNS, has dropped markedly. Some studies suggest that neurological involvement of infected patient occur with different frequency, depending on HIV subtype involved in the infection. Subtype C may have reduced neuroinvasive capacity, possibly due to its different primary conformation of HIV transactivating regulatory protein (Tat), involved in monocyte chemotaxis. This review focus on physiopathologic aspects of HIV infection in CNS and its correlation with HIV clades.

O sistema nervoso central (SNC) e o sistema imunológico são considerados os principais órgãos alvo na infecção pelo HIV. As manifestações neurológicas diretamente relacionadas ao HIV são meningites virais aguda e crônica, demência associada ao HIV, mielopatia vacuolar e envolvimento do sistema nervoso periférico. Mudanças no diagnóstico e sobrevida têm mudado o aspecto da infecção pelo HIV, não mais considerada uma doença fatal e sim crônica. Após HAART, a incidência da maioria das doenças oportunistas, incluindo aquelas que afetam o SNC, reduziu-se significativamente. Alguns estudos sugerem que o envolvimento de pacientes infectados ocorre com frequência diferente, dependendo do subtipo de HIV. O subtipo C apresenta uma capacidade reduzida de neuroinvasão, possivelmente devido a conformação primária da sua proteína reguladora da transativação (Tat), que perde sua capacidade quimiotáxica. Esta revisão aborda aspectos fisiopatológicos da infecção do HIV no SNC e subtipos de HIV.

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil / Enterovirus e família herpesviridae como agentes etiológicos de meningites linfomonocitárias, no sul do Brasil

Viral meningitis is a common infectious disease of the central nervous system (CNS) that occurs worldwide. The aim of this study was to identify the etiologic agent of lymphomonocytary meningitis in Curitiba, PR, Brazil. During the period of July 2005 to December 2006, 460 cerebrospinal fluid (CSF) samples with lymphomonocytary meningitis were analyzed by PCR methodologies. Fifty nine (12.8 percent) samples were positive. Enteroviruses was present in 49 (83 percent) samples and herpes virus family in 10 (17 percent), of these 6 (10 percent) herpes simplex virus, 1 (2 percent) Epstein Barr virus, 2 (3 percent) human herpes virus type 6 and 1 (2 percent) mixed infection of enterovirus and Epstein Barr virus. As conclusion enterovirus was the most frequent virus, with circulation during summer and was observed with higher frequency between 4 to 17 years of age. PCR methodology is an important method for rapid detection of RNA enterovirus and DNA herpesvirus in CSF.

A meningite viral é uma síndrome infecciosa comum do sistema nervoso central (SNC), que ocorre no mundo inteiro. O objetivo deste estudo foi identificar o agente etiológico de meningite linfomonocitária em Curitiba, PR, Brasil. Durante o período de julho de 2005 a dezembro de 2006, 460 amostras com meningite linfomonocitária foram analisadas por metodologias de PCR. Cinquenta e nove (12,8 por cento) amostras foram positivas. Enterovirus estava presente em 49 (83 por cento) amostras e herpes vírus em 10 (17 por cento), destas 6 (10 por cento) HSV, 1 (2 por cento) EBV, 2 (3 por cento) HHV- 6 e 1 (2 por cento) infecção mista de enterovírus e EBV. Conclui-se que o enterovirus foi o vírus mais frequente, com a circulação durante o verão. Houve maior número de amostras positivas entre 4 a 17 anos. A metodologia de PCR é um importante método para a detecção rápida de RNA de enterovirus e DNA do herpesvirus no LCR.

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.

Molecular epidemiology of HIV-1 clades in Southern Brazil

Human immunodeficiency virus (HIV) clades B and C account for more than 60 percent of the HIV-1 infections worldwide. In this paper, we describe the profiles of patients infected with subtypes of HIV-1 from the state of Paraná, Southern Brazil, and correlate them with demographic and epidemiological findings. A retrospective analysis of HIV cases reported from 1999-2007 was also performed. Data from 293 patients were reviewed and 245 were older than 13 (58 percent female). The distribution of clades was as follows: B 140 (57 percent), C 67 (23 percent), F 24 (10 percent) and mosaic or unique recombinant forms (URFs) 24 (10 percent). Of the 48 patients younger than 13 years of age (62.5 percent male), vertical transmission occurred in 46 and the distribution of clades was as follows: B 14 (29 percent), C 24 (50 percent), F 7 (15 percent) and URFs 6 (13 percent). There was no significant difference in mortality between HIV-1 subtypes. In both groups, patients infected with clade C tended to have higher rates of injection drug use exposure risk.

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.

Comparison of immunoperoxidase and immunofluorescence assays for pp65 cytomegalovirus antigen in immunocompromised patients

We compared the pp65 antigen detection by an in house method (immunoperoxidase assay) and by a commercial kit (immunofluorescence assay) available for cytomegalovirus infection diagnosis in immunocompromised patients. Sixty-four blood samples were analyzed in duplicate for both techniques. Eight-six percent of the samples had concordant qualitative results. The discordant results occurred more frequently in samples with low quantity of positive cells. There were no significant differences with qualitative and quantitative results of the methods.

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.