ABSTRACT
Introducción. Los inmunoensayos de detección rápida de antígenos (TRA) del SARS-CoV-2, son considerados adecuados para el diagnóstico en el punto de atención. El objetivo fue conocer la concordancia entre la reacción en cadena de la polimerasa en tiempo real con transcriptasa inversa (RT-qPCR, por su sigla en inglés) y los TRA en población pediátrica. Población y métodos. Se reclutaron todos los pacientes entre 1 mes y 17 años 11 meses de edad atendidos en la Unidad Febril de Urgencia de un hospital pediátrico entre el 11 de junio y el 3 de octubre de 2021. Se utilizó el TRA Panbio COVID-19 Ag® (Abbott Diagnostic) y, comométodo de referencia, la RT-qPCR (según el protocolo de los Centros para el Control y la Prevención de Enfermedades). Resultados. Se incluyeron 6491 pacientes. La prevalencia de COVID-19 fue del 2,8 %. El92,1 % de los sujetos presentaron síntomas. La sensibilidad, la especificidad y el índice kappa de concordancia para el TRA fueron del 71,0 %, 99,9 % y 0,813, respectivamente. El índice kappa yla sensibilidad del TRA fueron significativamentemayores en el grupo de 13 a 17 años (0,89 y 82,4 %,respectivamente) cuando se los comparó con los grupos de 0 a 5 y de 6 a 12 años. Esto podría deberse a la menor carga viral observada en los pacientes menores de 12 años. Conclusión. Si bien los TRA permiten acortar el tiempo de obtención de los resultados y mejorar la estrategia de aislamiento de pacientes con COVID-19, la sensibilidad en niños menores de 12 años o asintomáticos no se encontraría dentro de los rangos recomendados, sobre todo enperíodos de baja prevalencia de la enfermedad.
Introduction. Rapid antigen tests (RAgTs) for SARS-CoV-2 are considered adequate for diagnosis at the point of care. Our objective was to establish the agreement between reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and RAgTs in the pediatric population. Population and methods. All patients aged 1 month to 17 years and 11 months seen at the Emergency Fever Unit of a children's hospital between 6-11-2021 and 10-3-2021 were recruited. The Panbio COVID-19 Ag® test (Abbott Diagnostic) was compared to the reference method RT-qPCR (as per the protocol suggested by the United States Centers for Disease Control and Prevention). Results. A total of 6491 patients were included. The prevalence of COVID-19 was 2.8%. Symptoms were observed in 92.1%. Sensitivity, specificity, and the kappa index of agreement for the RAgT were 71.0%, 99.9%, and 0.813, respectively. The kappa index and the RAgT sensitivity were significantly higher in the group aged 1317 years (0.89 and 82.4%, respectively) compared to the groups aged 05 and 612 years. This may be due to the lower viral load observed in patients younger than 12 years. Conclusion. Although RAgTs shorten the time to result and improve the isolation strategy for COVID-19 patients, their sensitivity in children younger than 12 years or asymptomatic children is not within the recommended ranges, especially during periods of low disease prevalence.
Subject(s)
Humans , Infant , Child, Preschool , Child , Adolescent , COVID-19/diagnosis , Sensitivity and Specificity , Emergency Service, Hospital , COVID-19 Testing , SARS-CoV-2 , Hospitals, PediatricABSTRACT
Frenar la propagación de la enfermedad por el coronavirus 2019 (COVID-19, por su sigla en inglés) es fundamental, y se puede realizar mediante técnicas de detección rápidas y efectivas. El objetivo fue comparar la precisión diagnóstica de un test rápido de antígeno (TRAg,) con la reacción en cadena de polimerasa con retrotranscripción (RT-qPCR, por su sigla en inglés) y describir los umbrales de amplificación (Ct, por su sigla en inglés). Participaron niños de 1 mes a 11 años que tuvieran menos de 7 días de síntomas, sin resultado detectable en los últimos 90 días, e inmunocompetentes. Se incluyeron 1855 pacientes con una prevalencia de COVID-19 del 4,7 %. La sensibilidad global del TRAg fue del 60,2 % y su especificidad, del 99,8 %; en niños mayores de 5 años los valores fueron de 69,8 % y 99,8 %, respectivamente. Los valores de Ct de las muestras discordantes fueron más altos. En conclusión, la precisión diagnóstica muestra que TRAg tiene una especificidad similar a la RT-qPCR, pero una sensibilidad considerablemente menor, sobre todo en niños de menos de 5 años.
Stopping the spread of coronavirus disease 2019 (COVID-19) is critical and can be achieved through rapid and effective detection techniques. Our objective was to compare the diagnostic accuracy of rapid antigen tests (RAgT) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) and to describe amplification cycle thresholds (Cts). Participants were children aged 1 month to 11 years with symptoms for less than 7 days, who did not have a detectable result in the past 90 days, and were immunocompetent. A total of 1855 patients were included; the prevalence of COVID-19 was 4.7%. For the RAgT, overall sensitivity was 60.2% and specificity, 99.8%; in children older than 5 years, values were 69.8% and 99.8%, respectively. Ct values for discordant samples were higher. To conclude, the diagnostic accuracy indicated that the specificity of RAgT is similar to that of RT-qPCR, but its sensitivity is notably lower,especially in children younger than 5 years.
Subject(s)
Humans , Infant , Child, Preschool , Child , SARS-CoV-2 , COVID-19/diagnosis , Cross-Sectional Studies , Sensitivity and Specificity , Clinical Laboratory Techniques/methods , Real-Time Polymerase Chain Reaction , COVID-19 TestingABSTRACT
Introducción: en diciembre del año 2019 surgió en China una neumonía viral; el virus fue identificado como un coronavirus SARS-CoV-2, que se propagó rápidamente de tal manera que se convirtió en pandemia. La alta contagiosidad y la presencia de portadores asintomáticos dificultaron el diagnóstico de la infección y la toma de decisiones sanitarias. Objetivo: el objetivo de esta revisión bibliográfica es presentar y describir las principales técnicas utilizadas actualmente para el diagnóstico de COVID-19 y establecer su relación con los conocimientos de distintas disciplinas y tecnologías emergentes que confluyen en la Biotecnología bioquímico-farmacéutica orientada a la Salud humana. Metodología: se realizó una revisión de la bibliografía disponible en PubMed a partir de enero de 2020 sobre las pruebas diagnósticas que se encuentran actualmente en uso, en el ámbito sanitario, para la detección y seguimiento de la enfermedad COVID-19. También se realizaron búsquedas a través de Google y Google Académico para publicaciones de organismos de Salud en referencia a métodos diagnósticos. Resultados: se presenta una importante cantidad de pruebas diagnósticas, basadas en diferentes tecnologías, que desempeñan un papel clave en la pandemia de COVID-19. Algunas de ellas muy sofisticadas, como la secuenciación genómica de próxima generación, otras más estándar, pero igualmente robustas, como la reacción en cadena de la polimerasa (PCR). También otras adaptadas para el brote pandémico, como la amplificación isotérmica de ácidos nucleicos mediada por bucle. Todas las mencionadas se consideran de tipo molecular, pero también existen las pruebas serológicas, como ELISA, que incluyen ensayos en plasma o de tipo inmunológico. Estas sirven para detectar anticuerpos frente a la exposición al virus o antígenos en personas potencialmente infectadas. Conclusiones: los procesos de investigación y desarrollo biotecnológicos aplicados al diagnóstico y los conocimientos científicos previos permitieron una respuesta tanto nacional como internacional rápida y eficaz en medio de una inédita pandemia global. En esta revisión destacamos las principales técnicas, en qué estadio se deben usar y qué información nos aportan. (AU)
Introduction: in December 2019, a viral pneumonia emerged in China, identifying the virus as a SARS-CoV-2 coronavirus, which spread rapidly in such a way that it became a pandemic. The high contagiousness and the presence of asymptomatic carriers make difficulted to diagnose the infection and to make health decisions. Target: the objective of this review is to present and describe the main techniques currently used for the diagnosis of COVID-19, and to establish their relationship with the knowledge of different disciplines and emerging technologies that converge in biochemical-pharmaceutical biotechnology oriented to human health. Methodology: a review of the literature available in Pubmed from January 2020 on the diagnostic tests that are currently in use in the health field, for the detection and monitoring of COVID-19 disease, was carried out. Searches were also carried out through Google and Google Scholar for publications of Health organizations in reference to diagnostic methods. Results: a significant number of diagnostic tests are presented, based on different technologies, which play a key role in the COVID-19 pandemic. Some of them are very sophisticated, such as next-generation genomic sequencing, others more standard, but equally robust, such as polymerase chain reaction. Also others adapted for the pandemic outbreak such as loop-mediated isothermal amplification of nucleic acids. All of the aforementioned are considered molecular, but there are also serological tests, such as ELISA, which include plasma or immunological tests. These serve to detect antibodies against exposure to the virus or antigens in potentially infected people. Conclusions: biotechnological research and development processes applied to diagnosis and previous scientific knowledge allowed a rapid and effective national and international response in the midst of an unprecedented global pandemic. In this review we highlight the main techniques, at what stage they should be used and what information they provide us. (AU)
Subject(s)
Humans , Biotechnology/trends , COVID-19 Testing/methods , SARS-CoV-2/isolation & purification , COVID-19/diagnosis , Enzyme-Linked Immunosorbent Assay , Biomarkers , Sensitivity and Specificity , Diagnostic Techniques and Procedures , COVID-19 Nucleic Acid Testing , COVID-19 Serological TestingABSTRACT
Abstract Objectives: to characterize school-aged children, adolescents, and young people's profile and their associations with positive COVID-19 test results. Methods: an observational and descriptive study of secondary data from the COVID-19 Panel in Espírito Santo State in February to August 2020. People suspected of COVID-19, in the 0-19-years old age group, were included in order to assess clinical data and demographic and epidemiological factors associated with the disease. Results: in the study period, 27,351 COVID-19 notification were registered in children, adolescents, and young people. The highest COVID-19 test confirmation was found in Caucasians and were 5-14 years age group. It was also observed that headache was the symptom with the highest test confirmation. Infection in people with disabilities was more frequent in the confirmed cases. The confirmation of cases occurred in approximately 80% of the notified registrations and 0.3% of the confirmed cases, died. Conclusion: children with confirmed diagnosis for COVID-19 have lower mortality rates, even though many were asymptomatic. To control the chain of transmission and reduce morbidity and mortality rates, it was necessaryto conduct more comprehensive research and promote extensive testing in the population.
Resumo Objetivos: Caracterizar o perfil de crianças, adolescentes e jovens em idade escolar e associações com o resultado positivo do teste COVID-19. Métodos: estudo observacional e descritivo de dados secundários do Painel COVID-19, no Estado do Espírito Santo no período de fevereiro a agosto de 2020. Foram incluídas pessoas suspeitas de COVID-19, em faixas etárias de 0 a 19 anos, a fim de avaliar os dados clínicos e fatores demográficos e epidemiológicos associados ao agravo. Resultados: no período de estudo, foram considerados 27.351 registros de notificação da COVID-19 em crianças, adolescentes e jovens. As maiores chances de confirmação dos casos foram encontradas na faixa etária de 5 a 14 anos, em pessoas de raça/cor branca para COVID-19. Observouse que cefaleia foi o sintoma que apresentou maior chance de confirmação de teste. Já a infecção em pessoas deficientes foram mais frequentes nos casos confirmados. A confirmação dos casos se deu em aproximadamente 80% dos registros de notificação e do total confirmados 0,3% vieram a óbito. Conclusão: as crianças com diagnóstico confirmado para COVID-19 apresentam menor taxa de mortalidade, mesmo que muitas fossem assintomáticas. Para o controle da cadeia de transmissão e redução nas taxas de morbimortalidade, torna-se necessária a realização de pesquisas mais abrangentes e promoção da testagem ampla na população.
Subject(s)
Humans , Child , Adolescent , Adult , Indicators of Morbidity and Mortality , Disease Notification/statistics & numerical data , COVID-19 Testing/statistics & numerical data , COVID-19/prevention & control , COVID-19/epidemiology , Brazil/epidemiology , Demography , Epidemiology, Descriptive , Observational StudyABSTRACT
RESUMEN En el Perú, la pandemia de la COVID-19 ha evidenciado la utilidad de tener un sistema de vigilancia laboratorial estructurado y en funcionamiento desde hace 22 años, basado en la vigilancia de influenza; inicialmente en modalidad de unidades centinela, y después fortaleciéndose e innovándose, con recursos propios y con apoyo externo, para generar información de calidad. Se han implementado avances biotecnológicos para la confirmación diagnóstica e incrementado las capacidades de la red nacional de laboratorios, manteniendo la eficiencia, considerando las diversas y complejas realidades de los niveles regionales, y superando dificultades de comunicación y articulación entre instituciones. Resulta necesario consolidar este sistema, con trabajo colaborativo y coordinado entre sus componentes, impulsando su eficacia y oportunidad y promoviendo la vigilancia genómica de nuevos virus y variantes, como actualmente ocurre con el SARS-CoV-2.
ABSTRACT In Peru, the COVID-19 pandemic demonstrated the usefulness of having a structured laboratory surveillance system that has been operational for 22 years, based on influenza surveillance; initially in the form of sentinel units, and later strengthened and innovated, with its own resources and with external support, to provide quality information. Biotechnological advances have been implemented for diagnostic confirmation and the capacity of the national laboratory network has been expanded, maintaining efficiency, considering the diverse and complex realities of each region, and overcoming difficulties regarding communication and articulation between institutions. It is necessary to consolidate this system, with collaborative and coordinated work between its components, boosting its effectiveness and timeliness and promoting genomic surveillance of new viruses and variants, as is currently the case with SARS-CoV-2.
Subject(s)
Viruses , Epidemiologic Surveillance Services , Public Health Surveillance , SARS-CoV-2 , Influenza A virus , Influenza B virus , Health Surveillance , Molecular Diagnostic Techniques , Public Health Laboratory Services , National Health Systems , Epidemiological Monitoring , COVID-19 TestingABSTRACT
OBJETIVO: Investigar a relação entre a realização de testes para detectar COVID-19 e indícios de sofrimento psíquico, estresse e burnout entre profissionais de saúde da linha de frente da pandemia em um hospital geral. MÉTODOS: Estudo prospectivo de abordagem mista usando SRQ-20, PSS, OBI e entrevistas em profundidade em série de três entrevistas em 2020. RESULTADOS: Prevalências preocupantes de escores elevados de SRQ20, Burnout e Estresse Percebido ocorreram nas três entrevistas, e o registro de testes realizados foi crescente no período estudado, mas não houve associação entre desfechos e realização de testes para detectar COVID-19. Os temores de contrair a doença e de ser transmissor do vírus apareceram como os principais estressores para profissionais de saúde, mantidos apesar da realização de testes. CONCLUSÃO: Nesse grupo, testes realizados não se mostraram suficientes para modificar os efeitos psicossociais da atividade em linha de frente sobre profissionais de saúde.
OBJECTIVE: To investigate the relationship between the performance of tests to detect COVID-19 and signs of psychological distress, perceived stress and burnout among health professionals on the frontline of the pandemic in a general hospital. METHODS: Prospective mixed-approach study using SRQ-20, PSS, OBI and in-depth interviews in a series of 3 interviews throughout 2020. RESULTS: Worrying prevalences of high scores of SRQ20, Burnout and Perceived Stress occurred on the three interviews and the number of tests performed increased during the study period, but there was no association between outcomes and testing to detect COVID- 19. Fear of contracting the disease or being a transmitter of the virus appeared as the main stressors for health professionals, maintained despite testing. CONCLUSION: In this group, tests performed were not sufficient to modify the psychosocial effects of the front line activity on health professionals.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Young Adult , Burnout, Professional/psychology , Burnout, Professional/epidemiology , Health Personnel/psychology , Fear/psychology , Pandemics , COVID-19/transmission , Prevalence , Interviews as Topic , Prospective Studies , Psychological Distress , COVID-19 Testing , Hospitals, GeneralABSTRACT
INTRODUCCIÓN: El umbral de ciclo (en inglés cycle threshold-Ct) de la reacción de polimerasa en cadena en tiempo real con transcripción reversa (RT-qPCR) indica la concentración relativa de una secuencia de ARN; este valor se ha relacionado con la expresión de cuadros clínicos en infecciones virales. OBJETIVO: Determinar la correlación entre el valor Ct y la clasificación clínica de la COVID-19. MÉTODO: Se realizó un estudio transeccional correlacional; los valores Ct se obtuvieron mediante RT-qPCR dirigida al gen N del SARS-CoV-2 agrupándolos mediante un estimador robusto central y relacionándose con la clasificación clínica de la COVID-19. RESULTADOS: De los 718 casos incluidos en el estudio; 77,7% (558) fueron leves; 21,3% (153) moderados y 1% (7) graves. El valor Ct se agrupó en niveles: Ct bajo 18,83 - 30,10 y Ct alto > 30,10. Existió correlación significativa inversa débil (p = 0,002; rho de Spearman = -0,117) entre el valor Ct y la clasificación clínica. Las características: sexo, edad menor a 65 años, fiebre, escalofrío, diarrea, anosmia y sobrepeso-obesidad estuvieron asociadas al valor de Ct. CONCLUSIÓN: A menor valor Ct se espera una clasificación de mayor gravedad de la COVID-19; no obstante, debido a que la correlación es débil, su utilidad como predictor de gravedad es limitada.
BACKGROUND: The cycle threshold (Ct) of real-time reverse transcription PCR (RT-qPCR) indicates the relative concentration of an RNA sequence, this value has been related to clinical profile in viral infections. AIM: To determine the correlation between the Ct value and the clinical classification of COVID-19. METHOD: A correlational cross-sectional study was carried out, the Ct values were obtained by RT-qPCR directed to the N gene of SARS-CoV-2, grouping them by means of a central robust estimator and related to the clinical classification of COVID-19. RESULTS: Of the 718 cases included in the study; 77.7% (558) were mild; 21.3% (153) moderate and 1% (7) severe. The Ct value was grouped into levels: low Ct 18.83-30.10 and high Ct> 30.10. There was a weak inverse significant correlation (p = 0.002; Spearman's rho = -0.117) between the Ct value and the clinical classification. The characteristics: sex, age under 65 years, fever, chills, diarrhea, anosmia, and overweightobesity were associated with the Ct value. CONCLUSION: The lower the Ct value, a classification of greater severity of COVID-19 is expected, however, because the correlation is weak, its usefulness as a severity predictor is limited.
Subject(s)
Humans , Male , Female , COVID-19/diagnosis , Cross-Sectional Studies , Real-Time Polymerase Chain Reaction , COVID-19 Testing , SARS-CoV-2/geneticsABSTRACT
RESUMO: Introdução: apesar de ser o país de maior média de idade no mundo, o Japão tem se destacado no combate à pandemia da COVID-19 (do inglês Coronavirus Disease 2019) ao apresentar reduzidas taxas de contaminação pelo vírus e de mortalidade. Objetivo: discutir acerca das estratégias em saúde adotadas pelo Japão diante da pandemia da doença da COVID-19, bem como avaliar os dados sobre contaminação e mortalidade japoneses em comparação com os outros quatro países com maior média de idade do mundo (Itália, Alemanha, Portugal e Espanha) e o Brasil. Metodologia: para avaliação das estratégias em saúde japonesas foi realizada busca nas bases de dados: PubMed, Cochrane e Scielo, utilizando-se combinação dos termos "Japão", "covid", "coronavirus" e "sistemas de saúde", nos idiomas Inglês, Espanhol e Português. Os dados de infecção da COVID-19 foram extraídos do site Our World in Data, correspondendo ao período de 25 de janeiro de 2020 a 30 de julho de 2020. Resultados: dentre as medidas adotadas pelo país no enfrentamento à pandemia, destacam-se o diagnóstico e resposta precoces à infecção, o rastreamento de contatos, o diagnóstico precoce e disponibilidade de cuidados intensivos para pacientes graves e estímulo a medidas comportamentais de distanciamento. Dentre os países analisados, o Japão apresenta as menores taxas de contaminação e mortalidade em termos absolutos pela COVID-19. Conclusões: medidas de distanciamento social, diagnóstico e tratamento precoces parecem ter contribuído para o sucesso no combate à COVID-19 no Japão. No período estudado, em milhão de habitantes, o Japão teve 6,13 casos de Covid, enquanto o Brasil apresentou 218,26 casos. Já no número de mortes confirmadas pela doença, o primeiro teve uma taxa de 0,23 enquanto o segundo de 5,16 casos por milhão de habitantes. É possível, a partir do conhecimento dessas medidas, buscar mecanismos semelhantes ao traçar políticas de saúde no enfrentamento de pandemias em outros países. (AU)
ABSTRAC: Introduction: despite being the country with the highest average age globally, Japan has stood out in the fight against the COVID-19 (Coronavirus Disease 2019) pandemic by presenting low contamination rates by the virus and mortality. Objective: we aim to discuss the health strategies adopted by Japan in the face of the COVID-19 disease pandemic, as well as to evaluate data on Japanese contamination and mortality compared to the other four countries with the highest average age in the world (Italy, Germany, Portugal and Spain) and Brazil. Methodology: the search was carried out to evaluate Japanese health strategies by using the following databases: PubMed, Cochrane, and Scielo using a combination of the terms "Japan", "covid", "coronavirus" and "health systems" in English, Spanish and Portuguese. The COVID-19 infection data was extracted from the Our World in Data website, from January 25, 2020, to July 30, 2020. Results: Among the measures adopted by the country to face the pandemic, the early diagnosis and response to infection, contact tracing, early diagnosis and availability of intensive care for critically ill patients, and encouraging behavioral distancing measures stand out. Among the countries analyzed, Japan has the lowest rates of contamination and mortality in absolute terms by COVID-19. Conclusions: social distancing measures, early diagnosis, and treatment seem to have contributed to the success in combating COVID-19 in Japan. In the studied period, in a million inhabitants, Japan had 6.13 cases of covid while Brazil had 218.26 cases. As for the number of deaths confirmed by the disease, the first had a rate of 0.23 while the second had 5.16 cases per million inhabitants. Based on the knowledge of these measures, it is possible to seek similar mechanisms when designing health policies to face pandemics in other countries.(AU)
Subject(s)
Quality of Health Care , Health Systems/trends , Health Strategies , Early Diagnosis , Physical Distancing , COVID-19 Testing , COVID-19/prevention & control , Health Policy , Japan , Asian PeopleABSTRACT
Background: South Africa had over 4 million cases of coronavirus disease 2019 (COVID-19) infections and more than 1 million COVID-19-related deaths. Despite the devastating psychological impact of the COVID-19 pandemic, there is little qualitative, critical evaluation of government mental health services in this resource-limited setting. Aim: The authors describe the clinical service plan and response to the COVID-19 pandemic at a government psychiatric hospital. Setting: KwaZulu-Natal, South Africa. Methods: A descriptive narrative overview of the specialised psychiatric hospital's clinical response (April 2020 March 2021) to the COVID-19 pandemic was undertaken in the following domains: screening policy; testing and swabbing policy; staff training and monitoring; and restructuring the wards to accommodate mental health care users (MHCUs) with suspected cases of COVID-19. Results: The in-depth narrative reviews led to the introduction of staff training, routine COVID-19 reverse transcription polymerase chain reaction (RT-PCR) testing of all MHCUs, the creation of designated quarantine and isolation facilities and screening of physical health status of patients with COVID-19 prior to transfer being implemented to prevent an outbreak or increased morbidity or mortality. Conclusion: Implementing a service plan early which included staff training, screening and routine COVID-19 testing services for psychiatric admissions in a rapidly evolving environment with few additional resources was challenging. The absence of guidelines early in the pandemic that addressed the unique needs of a clinical psychiatric inpatient population is a noteworthy learning point. Contribution: The article highlights that the inpatient infrastructural requirements and clinical management protocols of acutely psychiatrically ill inpatients, in the context of infectious outbreaks, require dedicated task teams and bespoke policies.
Subject(s)
Humans , Male , Female , Mental Health , COVID-19 Testing , COVID-19 , Mental Health Services , Pandemics , InpatientsABSTRACT
The world has faced multiple waves of COVID-19 outbreaks, with more than 300 million cases and 5.5 million deaths officially reported globally as of Jan 8, 2022. Within the first year of the pandemic, there was hope that it would soon be under control, yet the pandemic sustains to be the world's priority health agenda. This brief communication provides emerging time-sensitive perspectives on the need for a tailored COVID-19 vaccination strategy in Ethiopia by reviewing studies and expert opinions. As of Jan 8, 2022, Ethiopia has reported 443,339 cases and 7,020 COVID-19-related deaths. Only 9,361,640 people (8%) of the Ethiopian population received at least one dose of the COVID-19 vaccine. While the short supply of vaccines is mentioned as a major bottleneck, the role of vaccine skepticism is largely overlooked, though the vaccine is the primary means to combat the emergence of new variants. Therefore, we recommend vaccine advocacy and awareness creation, planning for vaccine mandate for certain groups of the society, and targeted vaccination and economical use of the vaccines.
Subject(s)
Vaccination Coverage , COVID-19 Vaccines , COVID-19 Testing , COVID-19 , SARS-CoV-2ABSTRACT
ABSTRACT Objective: To describe the first COVID-19 pandemic at Casa Ondina Lobo, a philanthropic nursing home in São Paulo city, and the containment measures against the pandemic that proved to be effective. Methods: Several preventive measures were taken before and during the pandemic, with emphasis on universal testing by reverse transcription polymerase chain reaction for COVID-19. All residents and employees were tested twice in a D9 period. Results: Among the 62 residents and 55 employees, in both testing, eight residents and nine employees tested positive for COVID-19. Of 22% of employees and 75% of residents evolved asymptomatic, emphasizing the importance of universal testing for the detection and isolation of these cases. A quarter of residents evolved without any symptoms, however, with COVID-19 signs, reinforcing the importance of monitoring vital signs. The second testing did not detect any new cases among residents, demonstrating the effectiveness of the containment measures, however, it found four new cases among employees. This emphasized their role in COVID-19 outbreaks in nursing homes. Only one patient died, a 12.5% lethality among those known to be infected and a 1.6% mortality in the total population of residents were seen. Conclusion: The adoption of appropriate containment measures enabled to contain an COVID-19 pandemic in studied nursing home. Universal reverse transcription polymerase chain reaction testing for COVID-19 has proved to be particularly important and effective.
Subject(s)
Humans , COVID-19/prevention & control , Brazil/epidemiology , Pandemics/prevention & control , COVID-19 Testing , Nursing HomesABSTRACT
ABSTRACT Acute vulvar ulcer (Lipschütz's ulcer) is a rare lesion with local hyperimmunoreactivity triggered by infection, which is characterized by acute, painful, and necrotic ulcerations. This condition is usually found in non-sexually active adolescents, and it resolves spontaneously. We report a case of a 35-year-old woman who was diagnosed with COVID-19 who did not have severe symptoms, but had high levels of D-dimer for 9 days. The COVID-19 diagnosis was followed by the appearance of an acute, necrotic, extremely painful vulvar ulcer, although symptoms caused by COVID-19 had improved. We emphasize the importance of the differential diagnosis to exclude diseases such as Behçet's syndrome, Sexually Transmitted Infections, as well as the presence of viruses that generally trigger Lipschütz's ulcer, such as Epstein-Barr virus and cytomegalovirus. No treatment is usually necessary, however, in the present report due to the pain experienced by the patient, we successfully used oral prednisone.
Subject(s)
Humans , Female , Adolescent , Adult , Behcet Syndrome/complications , Behcet Syndrome/diagnosis , Behcet Syndrome/drug therapy , Epstein-Barr Virus Infections , COVID-19 , Ulcer/drug therapy , Herpesvirus 4, Human , COVID-19 Testing , SARS-CoV-2 , GenitaliaABSTRACT
O objetivo foi analisar as associações entre a percepção de risco de adoecimento por COVID-19 e os sintomas de depressão, ansiedade e estresse em profissionais atuantes em unidades de saúde. Estudo transversal com trabalhadores de diversas categorias profissionais que buscaram voluntariamente um dos primeiros Centros de Referência em Testagem de COVID-19 no Município do Rio de Janeiro, Brasil. Os trabalhadores foram convidados a responder a um questionário online entre maio e agosto de 2020. Foram utilizadas a escala Percepção de Risco de Adoecimento por COVID-19 e a Escala de Depressão, Ansiedade e Estresse (DASS-21). Foram estimados razão de chance (OR) e intervalo de 95% de confiança. Do total (N = 2.996), 81,5% eram mulheres com idade média de 40,7 anos. Cerca da metade apresentava grau leve, moderado ou severo de depressão, ansiedade ou estresse, sendo a frequência de trabalhadores com sintomas severos, respectivamente, 18,5%, 29,6% e 21,5%. Observou-se que as associações entre a percepção de risco e os sintomas de depressão, ansiedade e estresse foram mais fortes à medida que aumentava a classificação de gravidade de cada sintoma. Os trabalhadores com alta percepção de risco de adoecimento por COVID-19 apresentaram OR mais elevadas para sintomas severos de depressão (OR = 4,67), ansiedade (OR = 4,35) e estresse (OR = 4,97). Os achados apontam a demanda por medidas de proteção à saúde dos trabalhadores, que não devem se restringir aos equipamentos de proteção individual. É essencial que os gestores promovam espaços coletivos de discussão e ações que favoreçam a recuperação dos trabalhadores em contexto pandêmico de longa duração.
The objective was to analyze associations between perceived risk from COVID-19 and symptoms of depression, anxiety, and stress among workers in healthcare units. This was a cross-sectional study of workers from different professions who appeared voluntarily at one of the first COVID-19 Testing Centers in the municipality of Rio de Janeiro, Brazil. The workers were invited to answer an online questionnaire from May to August 2020. The COVID-19 Risk Perception Scale and the Depression, Anxiety, and Stress Scale (DASS-21) were used. Odds ratios (OR) and 95% confidence intervals were estimated. Of the total sample (N = 2,996), 81.5% were women, and mean age was 40.7 years. About half presented mild, moderate, or severe depression, anxiety, or stress, and the rates for workers with severe symptoms were 18.5%, 29.6%, and 21.5%, respectively. The associations between perceived risk and symptoms of depression, anxiety, and stress increased with the increase in each symptom's severity. Workers with higher perceived risk from COVID-19 showed higher OR for severe symptoms of depression (OR = 4.67), anxiety (OR = 4.35), and stress (OR = 4.97). The findings point to the demand for measures to protect workers' health and that should not be limited to personal protective equipment. It is essential for health system administrators to promote collective spaces for discussion and actions to favor workers' recovery in the context of a prolonged pandemic.
El objetivo fue analizar las asociaciones entre la percepción de riesgo de enfermedad por COVID-19 y los síntomas de depresión, ansiedad y estrés en profesionales activos en unidades de salud. Estudio transversal con trabajadores de diversas categorías profesionales que buscaron voluntariamente uno de los primeros Centros de Referencia en Tests de COVID-19 en el municipio de Río de Janeiro, Brasil. Los trabajadores fueron invitados a responder a un cuestionario online entre mayo y agosto de 2020. Se utilizaron la escala Percepción de Riesgo de Enfermedad por COVID-19 y la Escala de Depresión, Ansiedad y Estrés (DASS-21). Se estimaron razón de oportunidad (OR) e intervalo de 95% de confianza. Del total (N = 2.996), un 81,5% eran mujeres con una edad media de 40,7 años. Cerca de la mitad presentaba grado leve, moderado o severo de depresión, ansiedad o estrés, siendo la frecuencia de trabajadores con síntomas severos, respectivamente, 18,5%, 29,6% y 21,5%. Se observó que las asociaciones entre la percepción de riesgo y los síntomas de depresión, ansiedad y estrés fueron más fuertes a medida que aumentaba la clasificación de la gravedad de cada síntoma. Los trabajadores con alta percepción de riesgo de enfermedad por COVID-19 presentaron OR más elevadas para síntomas severos de depresión (OR = 4,67), ansiedad (OR = 4,35) y estrés (OR = 4,97). Los resultados apuntan la demanda de medidas de protección a la salud de los trabajadores, que no se deben restringir a equipamientos de protección individual. Es esencial que los gestores promuevan espacios colectivos de discusión y acciones que favorezcan la recuperación de los trabajadores en un contexto pandémico de larga duración.
Subject(s)
Humans , Female , Adult , Depression/epidemiology , COVID-19 , Anxiety/epidemiology , Stress, Psychological/diagnosis , Stress, Psychological/epidemiology , Brazil/epidemiology , Cross-Sectional Studies , Delivery of Health Care , COVID-19 TestingABSTRACT
Point-of-care serological tests for SARS-CoV-2 have been used for COVID-19 diagnosis. However, their accuracy over time regarding the onset of symptoms is not fully understood. We aimed to assess the accuracy of a point-of-care lateral flow immunoassay (LFI). Subjects, aged over 18 years, presenting clinical symptoms suggestive of acute SARS-CoV-2 infection were tested once by both nasopharyngeal and oropharyngeal RT-PCR and LFI. The accuracy of LFI was assessed in periodic intervals of three days in relation to the onset of symptoms. The optimal cut-off point was defined as the number of days required to achieve the best sensitivity and specificity. This cut-off point was also used to compare LFI accuracy according to participants' status: outpatient or hospitalized. In total, 959 patients were included, 379 (39.52%) tested positive for SARS-CoV-2 with RT-PCR, and 272 (28.36%) tested positive with LFI. LFI best performance was achieved after 10 days of the onset of symptoms, with sensitivity and specificity of 84.9% (95%CI: 79.8-89.1) and 94.4% (95%CI: 91.0-96.8), respectively. Although the specificity was similar (94.6% vs. 88.9%, p = 0.051), the sensitivity was higher in hospitalized patients than in outpatients (91.7% vs. 82.1%, p = 0.032) after 10 days of the onset of symptoms. Best sensitivity of point-of-care LFI was found 10 days after the onset of symptoms which may limit its use in acute care. Specificity remained high regardless of the number of days since the onset of symptoms.
Os testes sorológicos no local de atendimento (point-of-care) para a infecção pelo SARS-CoV-2 têm sidos utilizados para o diagnóstico da COVID-19. Entretanto, não está plenamente elucidada a acurácia dos testes ao longo do tempo em relação ao início dos sintomas. Nosso objetivo foi de avaliar a acurácia, no local de atendimento, do imunoensaio de fluxo lateral (LFI). Pacientes com ≥ 18 anos de idade que apresentavam sintomas clínicos sugestivos de infecção aguda pelo SARS-CoV-2 foram testados uma vez com RT-PCR da nasofaringe e orofaringe, além do LFI. A acurácia do LFI foi avaliada com intervalos periódicos de 3 dias a partir do início dos sintomas. O ponto de corte ótimo foi definido como o número necessário de dias para atingir a melhor sensibilidade e especificidade. Esse ponto foi utilizado também para comparar a acurácia do LFI de acordo com a situação do paciente (ambulatorial ou hospitalizado). Foram incluídos 959 pacientes, dos quais 379 (39,52%) testaram positivos para SARS-CoV-2 pelo RT-PCR e 272 (28,36%) pelo LFI. Foi atingido o melhor desempenho para o LFI com 10 dias a partir do início dos sintomas, com sensibilidade e especificidade de 84,9% (IC95%: 79,8-89,1) e 94,4% (IC95%: 91,0-96,8), respectivamente. Embora a especificidade não tenha sido diferente entre os grupos de pacientes (94,6% vs. 88,9%, p = 0,051), a sensibilidade foi mais alta nos pacientes hospitalizados que nos ambulatoriais (91,7% vs. 82,1%, p = 0,032) no dia 10 depois do início dos sintomas. A melhor sensibilidade do LFI no local de atendimento ocorre 10 dias depois do início dos sintomas, o que pode limitar seu uso no atendimento agudo. A especificidade permanece alta, independentemente do número de dias desde o início dos sintomas.
Los puestos de atención para pruebas serológicas del SARS-CoV-2 han sido usado para la diagnosis de la COVID-19. No obstante, su precisión a lo largo del tiempo, en lo que respecta a la aparición de los síntomas, no se ha comprendido completamente. Nuestro objetivo fue evaluar la precisión de un puesto de atención de inmunoanálisis de flujo lateral (LFI). Se hizo pruebas a individuos ≥ 18 años, presentando síntomas clínicos compatibles con una infección aguda de SARS-CoV-2, tanto vía nasofaríngea y orofaríngea RT-PCR, como LFI. La precisión de LFI fue evaluada en intervalos periódicos de 3 días con respecto a la aparición de los síntomas. El punto óptimo de corte se definió como el número de días requerido para alcanzar la mejor sensibilidad y especificidad. Este punto también se usó para comparar la precisión del LFI, según el estatus de los participantes: ambulatorios u hospitalizados. Se incluyeron a 959 pacientes, 379 (39,52%) dieron positivo en las pruebas de SARS-CoV-2 RT-PCR, y 272 (28,36%) fueron positivos en los LFI. Se alcanzó el mejor rendimiento de los LFI tras 10 días de la aparición de los síntomas, con una sensibilidad y especificidad de un 84,9% (IC95%: 79,8-89,1) y 94,4% (IC95%: 91,0-96,8), respectivamente. A pesar de que la especificidad no fue diferente (94,6% vs. 88,9%, p = 0,051), la sensibilidad fue mayor en pacientes hospitalizados que en los ambulatorios (91,7% vs. 82,1%, p = 0,032) tras 10 días desde la aparición de los síntomas. La mejor sensibilidad LFI del puesto de cuidado se produce tras 10 días de la aparición de los síntomas, lo que quizás limite su uso en el cuidado de urgencias. La especificidad permanece alta independientemente del número de días desde la aparición de los síntomas.
Subject(s)
Humans , Adult , SARS-CoV-2 , COVID-19 , Brazil , Sensitivity and Specificity , COVID-19 Testing , Middle AgedABSTRACT
RESUMO Inicialmente, gestantes e puérperas não foram associadas a risco maior de morbimortalidade relacionada ao Sars-Cov-2. Contudo, a evolução da pandemia tem mostrado um quadro diferente com aumento do acometimento e agravamento dos casos repercutindo em crescentes complicações materno-fetais, especialmente no terceiro trimestre. Objetivos: Este estudo tem como objetivo apresentar dados preliminares de incidência na população de gestantes, atendidas em Hospital terciário do Sul do Brasil, referência no atendimento COVID, obtidos no primeiro ano de pandemia. Resultados: entre março de 2020 e março de 2021, 1801 gestantes foram admitidas no Centro Obstétrico. Destas, 128 (7,10%) referiram alguma queixa referente à síndrome gripal da infecção por coronavírus, sendo que 46,09% (59 gestantes) apresentaram positividade (contaminação pelo Sars-Cov-2) no teste RT-PCR, com variação sazonal. Em 2020, o pico de positividade foi em junho e julho (38,46% e 54,54% respectivamente), porém o pico mais evidente ocorreu em fevereiro (83,33%) e março de 2021 (78.9%) de positividade viral, entre as gestantes sintomáticas. No primeiro ano da pandemia, 16 gestantes internaram em centro de tratamento intensivo, correspondendo a 27,1% das sintomáticas e infectadas com Sars-Cov-2. Conclusões: Importante destacar a elevação significativa de casos em março de 2021 e a importância de conhecermos a evolução da COVID-19 na gestação, uma vez que entre gestantes sintomáticas com testes positivos, 22% necessitaram de CTI. O comportamento viral pode ser diferente, conforme a população atendida, estes dados podem ser fundamentais no planejamento de estratégias de saúde e de atendimento às mulheres no ciclo gravídico-puerperal.
ABSTRACT Initially, pregnant and postpartum women were not associated with an increased risk of SARS-Cov-2-related morbidity and mortality. However, the evolution of the pandemic has shown a different picture with an increase in the number and aggravation of cases, resulting in increasing maternal-fetal complications, especially in the third trimester. Objectives: This study aims to present preliminary incidence data in the population of pregnant women treated at a tertiary Hospital in South Brazil, a reference in COVID care, obtained in the first year of the pandemic. Results: Between March 2020 and March 2021, 1801 pregnant women were admitted to the Obstetric Center. Of these, 128 (7.10%) reported some complaint regarding the flu-like syndrome of coronavirus infection, and 46.09% (59 pregnant women) were positive for Sars-Cov-2 in the RT-PCR test, with seasonal variation. In 2020 the peak of positivity was in June and July (38.46% and 54.54%, respectively), but the most evident peak of viral positivity occurred in February (83.33%) and March 2021 (78.9%) among symptomatic pregnant women. In the first year of the pandemic, 16 pregnant women were admitted to an intensive care unit, corresponding to 27.1% of symptomatic women infected with Sars-Cov-2. Conclusions: It is important to highlight the significant increase in cases in March 2021 and the importance of knowing the evolution of COVID-19 during pregnancy, since among symptomatic pregnant women with positive tests, 22% required ICU. Viral behavior can be different, depending on the population served, these data can be fundamental in the planning of health and care strategies for women in the pregnancy-puerperal cycle.
Subject(s)
Humans , Severe acute respiratory syndrome-related coronavirus , COVID-19 Testing , COVID-19ABSTRACT
RESUMEN Se validó y evaluó un método de RT-PCR en tiempo real usando cebadores y sondas específicas para los genes RdRP de SARS-CoV-2 y GAPDH de humanos; este último fue usado como control endógeno. Se evaluó la especificidad y sensibilidad; además, se evaluó otros parámetros como la robustez, la repetibilidad, reproducibilidad, comparabilidad y el límite de detección. La sensibilidad, especificidad, los valores predictivos positivo y negativo, la robustez, comparabilidad y la repetibilidad-reproducibilidad de la prueba de RT-PCR en tiempo real dúplex fue de 100%, con un límite de detección de 100 copias/µL, de acuerdo con los criterios de aceptación establecidos para validación del protocolo. Esta prueba estandarizada es una buena alternativa para el diagnóstico de COVID-19; además, la prueba fue aplicada de manera exitosa en personas sospechosas de la enfermedad permitiendo controlar el número de falsos negativos.
ABSTRACT The present work validated and evaluated a duplex real-time RT-PCR using specific primers and probes for genes RdRp from SARS-CoV-2 and GAPDH from humans; the latter was used as an endogenous control in all reactions. We evaluated the specificity, the sensitivity, the robustness, the reproducibility, the repeatability, the comparability, and the limit of detection. The predictive positive and negative values (PPV and PNV, respectively) and all the parameters evaluated using our duplex real-time RT-PCR was 100%. The detection limit was 100 copies/µL according to the acceptance criteria established for the validation of this protocol. Our duplex real-time RT-PCR demonstrated to be a good alternative for the diagnosis of COVID-19; in addition, this PCR was used adequately in suspicion of COVID-19, allowing it to control the number of false-negatives.
Subject(s)
Validation Study , Molecular Diagnostic Techniques , SARS-CoV-2 , COVID-19 Testing , COVID-19ABSTRACT
Contar con métodos diagnósticos que reúnan ciertos atributos es vital para guiar las decisiones sanitarias, el contexto actual lo amerita. Con el objetivo de validar la capacidad de dos pruebas rápidas para detectar anticuerpos debido a la infección por SARS-CoV-2 en la Isla de la Juventud, Cuba, de abril a mayo de 2020, se realizó un estudio descriptivo de corte transversal de evaluación de pruebas rápidas: Wondfo (SARS-CoV-2 antibody test) y Lungene covid-19 IgG/IgM rapid test, en comparación con la prueba de reacción en cadena de la polimerasa en tiempo real. Se construyeron dos muestras homogéneas de 250 cada una, determinándose indicadores de validación. Se obtuvieron valores de sensibilidad de 6,6 por ciento y 8,3 por ciento respectivamente para cada prueba, mientras que la especificidad resultó superior para Wondfo (95 por ciento). Los valores predictivos positivos resultaron muy bajos, los negativos adecuados, superior en Lungene con el 94,8 por ciento. Los valores de razón de verosimilitud fueron clasificados como inútiles. En diferentes escenarios en cuanto a casos sintomáticos, se alcanzó sensibilidad del 50 por ciento en intervalo de 1 a 7 días para Wondfo. El área bajo la curva ROC para Wondfo fue 0,50 (IC95 por ciento=0,46-0,55) y para Lungene 0,46 (IC95 por ciento=0,38-0,55). El índice kappa para Wondfo fue de 0,025 y 0,010 para Lungene. Las pruebas rápidas exploradas mostraron muy baja sensibilidad, valor predictivo positivo y razón de verosimilitud inadecuada. La validez global de las pruebas no demostró un buen desempeño diagnóstico, marcado por el valor del área bajo la curva ROC(AU)
Having diagnostic methods that meet certain attributes is vital to guide health decisions, the current context warrants it. In order to validate the capacity of two rapid tests to detect antibodies due to SARS-CoV-2 infection in Isla de la Juventud, Cuba, from April to May 2020, a descriptive cross-sectional evaluation study was carried out. Rapid tests: Wondfo (SARS-CoV-2 antibody test) and Lungene covid-19 IgG/IgM, were compared to the real-time polymerase chain reaction test. Two homogeneous samples of 250 each were constructed, determining validation indicators. Sensitivity values of 6.6 percent and 8.3 percent respectively were obtained for each test, while the specificity was higher for Wondfo (95 percent). The positive predictive values were very low, the negative ones were adequate, higher in Lungene with 94.8 percent. Likelihood ratio values were classified as useless. In different scenarios in terms of symptomatic cases, sensitivity of 50 percent was reached in an interval of 1 to 7 days for Wondfo. The area under the ROC curve for Wondfo was 0.50 (95 percent CI = 0.46-0.55) and 0.46 for Lungene (95 percent CI = 0.38-0.55). The kappa index for Wondfo was 0.025 and 0.010 for Lungene. The rapid tests explored showed very low sensitivity, positive predictive value, and inadequate likelihood ratio. The global validity of the tests did not demonstrate a good diagnostic performance, marked by the value of the area under the ROC curve. The degree of agreement was poor(AU)
Subject(s)
Humans , Validation Studies as Topic , COVID-19 Testing/methods , SARS-CoV-2 , COVID-19 , Enzyme-Linked Immunosorbent Assay/methods , Epidemiology, Descriptive , Cross-Sectional Studies , CubaABSTRACT
Resumen Los pacientes con cáncer y COVID-19 tienen más complicaciones que la población general. Comunicamos una cohorte de 74 pacientes con cáncer y COVID-19 internados en una institución on cológica. El 87.8% tenía diagnóstico de tumores sólidos y 12.2% oncohematológicos. Entre los tumores sólidos, el 61.5% presentó enfermedad metastásica. El 78.3% (N = 58) tenía infiltrados pulmonares al diagnóstico de COVID-19. La infección fue intrahospitalaria en 20 pacientes. Habían recibido tratamiento antineoplásico den tro de los 30 días anteriores al diagnóstico 39 pacientes (52.7%); uno se encontraba recibiendo radioterapia. Veinticuatro pacientes (32.4%) se derivaron a terapia intensiva (UTI) y 18 (75%) de ellos requirieron asistencia respiratoria mecánica (ARM). La mortalidad general durante la internación fue 32.4% (N = 24). La mortalidad en UTI fue 62.5% (N = 15). La mortalidad en ARM fue 72.2% (N = 13). La edad, recuento de neutrófilos, índice neutrófilo/linfocito, dímero D, ferritina, tabaquismo y haber adquirido la infección durante la internación resultaron estadísticamente significativos en el análisis univariado para mortalidad. No hallamos diferencias en mortalidad por estadio de enfermedad, en los pacientes con tumores sólidos, ni por haber recibido tratamiento antineoplá sico dentro de los 30 días del diagnóstico de COVID-19. En el análisis multivariado con el modelo de regresión logística, solo la edad y el tabaquismo fueron predictores de mortalidad. Los odds ratios (IC 95) ajustados para la edad ≥65 años y el tabaquismo fueron 8.87 (1.35-58.02) y 8.64 (1.32-56.64), respectivamente. Este trabajo puede resultar de utilidad para instituciones polivalentes que asistan pacientes oncológicos durante la pandemia.
Abstract Cancer patients are exposed to more complications from COVID-19 than non-cancer patients. We report a cohort of 74 cancer patients (87.8% with solid neoplasia and 12.2% with hematological diseases) with COVID-19 infection admitted to a tertiary medical cancer center in Argentina. Pulmonary infiltrates were diagnosed at admission in 78.3% (N = 58) of the cases. COVID-19 infection was hospital-acquired in 20 (27.0%) patients. Thirty-nine patients (52.7%) received anticancer therapy within the 30 days prior to COVID-19 diagnosis; one was on radiation therapy. Twenty-four (32.4%) patients were admitted in the intensive care unit (ICU) and 18 (75.0%) required mechanical ventilation. All cause in-hospital mortality was 32.4% (N = 24) and ICU mortality was 62.5% (N = 15). Mortality under me chanical ventilation was 72.2% (N = 13). In the univariate analysis age, neutrophil count, neutrophil/lymphocyte index, D-dimer, ferritin, smoking, and nosocomial acquired infection were associated with in-hospital mortality. There were no statistically significant differences in mortality related to disease stage for solid tumors, neither cancer treatment within 30 days of COVID-19 diagnosis. Age and smoking were associated with mortality in the multivariate analysis. The adjusted odds ratios (95 CI) for age ≥ 65 years and smoking were 8.87 (1.35-58.02) and 8.64 (1.32 - 56.64), respectively. Our experience can be useful for other institutions that assist cancer patients during the pandemic.