Made available in DSpace on 2014-08-13T14:50:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-06-21Bitstream added on 2014-08-13T18:00:42Z : No. of bitstreams: 1 000756392.pdf: 1529746 bytes, checksum: c695f204ec592597ec7f94c736d1b9ff (MD5) / O Coronavírus felino (FCoV) é um membro da ordem Nidovirales, família Coronaviridae, gênero Alphacoronavírus, que é responsável por causar a peritonite infecciosa felina (PIF). Dentre os testes sorológicos utilizados no auxílio ao diagnóstico da doença, o método mais sensível e recomendado é o ELISA. O FCoV apresenta características que dificultam seu cultivo celular, sendo necessário o uso de antígenos recombinantes no ELISA. A proteína de nucleocapsídeo (N) do FCoV apresenta vantagens no seu uso como antígeno recombinante em testes sorológicos, por ser conservada, altamente imunogênica, e ser uma das mais produzidas durante a infecção viral. O objetivo deste trabalho foi padronizar e aplicar o ELISA indireto com uso de proteína N recombinante para detectar anticorpos anti-FCoV em soros de gatos naturalmente infectados. Um fragmento de rim de um felino que veio a óbito com PIF foi utilizado para extração de RNA viral, amplificação pela RT-PCR e sequenciamento da proteína N. A sequência da proteína N foi sintetizada com códons otimizados para expressão em Escherichia coli e inserida no plasmídeo pGS21a. Após sua produção, a proteína foi purificada. Foi realizado o Western blott com soro de gato naturalmente infectado para a confirmação da reatividade da proteína. Na padronização do ELISA indireto foi determinada o tipo de bloqueio, concentração de antígeno e diluição dos soros. Trezentos e oitenta e duas amostras de soros felinos foram testadas. Os resultados obtidos mostraram que a proteína produzida apresentou ótima reatividade no ELISA indireto. Foram encontradas diferenças de A450 entre os soros testados, onde se demonstrou uma alta capacidade de discriminação com A450 de 0,1 a 1,8, mostrando que o teste é quantitativo. O ELISA indireto padronizado para detecção de anticorpos anti-FCoV utilizando a proteína N recombinante é um método eficiente e pode ser aplicado em amostras de campo / The Feline Coronavírus (FCoV) is a member of the Nidovirales order, Coronaviridae family, Alphacoronavírus genus, which is responsible for causing Feline Infectious Peritonitis (FIP). Among the serological tests used in the disease diagnosis, ELISA is the most sensitive and recommended assay. FCoV has characteristics that difficult their cell culture growing, requiring the use of recombinant antigens in the ELISA. The FCoV nucleocapsid protein (N) is conserved, highly immunogenic and is the most protein produced during viral infection, being a great recombinant antigen in serological tests. The aim of this study was to standardize and apply the indirect ELISA using recombinant N protein to detect anti-FCoV antibodies in sera from naturally infected cats. A kidney fragment of died feline FIP positive was used for extraction of viral RNA, amplification by RT-PCR and sequencing of the protein N. The N protein sequence was synthesized with optimized codons for expression in Escherichia coli and inserted into pGS21a plasmid. The protein was produced, purified and the Western blotting was performed with sera from naturally infected cat to confirm the reactivity of the protein. To indirect ELISA standardization, the antigen concentration, serum dilution and the blocking reagent were determined. Three hundred eighty-two cat’s sera were tested. The obtained results showed that the protein presented good reactivity in indirect ELISA. The A450 differences from 0.1 to 1.8 showed the quantitative feature of the assay. The standardized indirect ELISA for detection of anti-FCoV using recombinant N protein is efficient and can be applied in field samples

Orientador: João Pessoa Araújo Junior / Banca: Luciane Alarcão Dias Melício / Banca: Marcelo de Souza Zanutto / Resumo: O Coronavírus felino (FCoV) é um membro da ordem Nidovirales, família Coronaviridae, gênero Alphacoronavírus, que é responsável por causar a peritonite infecciosa felina (PIF). Dentre os testes sorológicos utilizados no auxílio ao diagnóstico da doença, o método mais sensível e recomendado é o ELISA. O FCoV apresenta características que dificultam seu cultivo celular, sendo necessário o uso de antígenos recombinantes no ELISA. A proteína de nucleocapsídeo (N) do FCoV apresenta vantagens no seu uso como antígeno recombinante em testes sorológicos, por ser conservada, altamente imunogênica, e ser uma das mais produzidas durante a infecção viral. O objetivo deste trabalho foi padronizar e aplicar o ELISA indireto com uso de proteína N recombinante para detectar anticorpos anti-FCoV em soros de gatos naturalmente infectados. Um fragmento de rim de um felino que veio a óbito com PIF foi utilizado para extração de RNA viral, amplificação pela RT-PCR e sequenciamento da proteína N. A sequência da proteína N foi sintetizada com códons otimizados para expressão em Escherichia coli e inserida no plasmídeo pGS21a. Após sua produção, a proteína foi purificada. Foi realizado o Western blott com soro de gato naturalmente infectado para a confirmação da reatividade da proteína. Na padronização do ELISA indireto foi determinada o tipo de bloqueio, concentração de antígeno e diluição dos soros. Trezentos e oitenta e duas amostras de soros felinos foram testadas. Os resultados obtidos mostraram que a proteína produzida apresentou ótima reatividade no ELISA indireto. Foram encontradas diferenças de A450 entre os soros testados, onde se demonstrou uma alta capacidade de discriminação com A450 de 0,1 a 1,8, mostrando que o teste é quantitativo. O ELISA indireto padronizado para detecção de anticorpos anti-FCoV utilizando a proteína N recombinante é um método eficiente e pode ser aplicado em amostras de campo / Abstract: The Feline Coronavírus (FCoV) is a member of the Nidovirales order, Coronaviridae family, Alphacoronavírus genus, which is responsible for causing Feline Infectious Peritonitis (FIP). Among the serological tests used in the disease diagnosis, ELISA is the most sensitive and recommended assay. FCoV has characteristics that difficult their cell culture growing, requiring the use of recombinant antigens in the ELISA. The FCoV nucleocapsid protein (N) is conserved, highly immunogenic and is the most protein produced during viral infection, being a great recombinant antigen in serological tests. The aim of this study was to standardize and apply the indirect ELISA using recombinant N protein to detect anti-FCoV antibodies in sera from naturally infected cats. A kidney fragment of died feline FIP positive was used for extraction of viral RNA, amplification by RT-PCR and sequencing of the protein N. The N protein sequence was synthesized with optimized codons for expression in Escherichia coli and inserted into pGS21a plasmid. The protein was produced, purified and the Western blotting was performed with sera from naturally infected cat to confirm the reactivity of the protein. To indirect ELISA standardization, the antigen concentration, serum dilution and the blocking reagent were determined. Three hundred eighty-two cat's sera were tested. The obtained results showed that the protein presented good reactivity in indirect ELISA. The A450 differences from 0.1 to 1.8 showed the quantitative feature of the assay. The standardized indirect ELISA for detection of anti-FCoV using recombinant N protein is efficient and can be applied in field samples / Mestre

A finales de 2020 se aprobaron las vacunas desarrolladas en el mundo occidental contra el virus SARS-CoV-2, que ya están siendo inoculadas, conjuntamente con vacunas chinas y rusas. Mientras tanto, estamos en una segunda oleada de la enfermedad y el nuevo coronavirus se ha ido transformando para permitirse una mejor propagación, alojamiento y replicación en el ser humano. La enfermedad se manifiesta ahora con nueva sintomatología, mayor contagio, inclemencia y variación en el número de fallecimientos. La infección de la gestante por coronavirus se está presentando con severidad y consecuencias materno perinatales. Ya se inició la vacunación en gestantes y madres lactantes, previa conversación con su ginecólogo sobre los riesgos y beneficios. Este artículo ofrece un breve rel los acontecimientos que tuvieron lugar durante la transición de 2020 a 2021. / In late 2020, vaccines developed in the Western world against the SARS-CoV-2 virus were approved and are currently being inoculated, together with Chinese and Russian vaccines. In the meantime, we are in a second wave of the disease and the new coronavirus has been transforming to allow for better propagation, harboring and replication in humans. The disease now manifests itself with new symptoms, greater contagiousness, severity and variation in the number of deaths. Coronavirus infection of pregnant women is occurring with harshness and maternal and perinatal consequences. Vaccination has been initiated in pregnant women and nursing mothers, after discussion with their gynecologists about risks and benefits. This article provides a b ok place during the transition from 2020 to 2021.

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Introducción. El COVID-19 y la gestación es una nueva intercurrencia en la valoración de riesgos para la atención de la gestante. Desde el inicio de la pandemia en el país, los casos han ido en aumento. El primer caso atendido en el Hospital San Bartolomé fue el 12 de abril. Desde los primeros reportes de gestantes COVID+ en China, a la fecha, se tiene cada vez mayor información, siendo importante para fines del manejo de la gestante COVID+ que se conozca su epidemiología y los resultados perinatales. Objetivo. Determinar la epidemiología y resultados materno perinatales de COVID-19 en las gestantes del Hospital Nacional Docente Madre Niño San Bartolomé, Lima, Perú. Método. Estudio observacional de corte transversal, en los meses de abril a julio 2020. Se incluyó a todas las gestantes que llegaron a la emergencia obstétrica del Hospital San Bartolomé, a las cuales se les realizó una prueba de inmunocromatografía para IgM/IgG, para determinar la seroprevalencia de COVID-19. Las variables obstétricas y perinatales fueron recolectadas en una ficha de datos al ingreso a la emergencia. Resultados. Se realizó prueba rápida para SARS-CoV-19 a 345 gestantes que se hospitalizaron para atención de parto. La edad promedio fue 27 años, con 10% de adolescentes y 16% de mayores de 35 años; 60% tenía 2 a 4 embarazos, 38% de los partos fue vaginal, 15% de ellos pretérmino; 1,2% de las gestantes fue sintomática y 0,2% ingresó a la unidad de cuidados intensivos. El 61% de los recién nacidos pesó entre 2 500 y 3 500 g, 53% fue sexo masculino, 94% tuvo Apgar mayor de 7 al minuto, 3,3% con hisopado positivo dentro de las primeras 24 horas. Se presentaron 3% de óbitos. El 48% de las gestantes provino del Cono Norte de la ciudad de Lima. Conclusiones. Casi 100% de las gestantes fue asintomática y solo 0,2% tuvo complicaciones respiratorias. La culminación del parto por vía cesárea fue baja en relación a otras publicaciones; el motivo de cesárea fue por indicación obstétrica. Escaso número de neonatos tuvo hisopado positivo. Hubo mayor incidencia de óbitos en julio 2020. El mayor porcentaje de pacientes provino del Cono Norte de Lima. / Introduction: COVID-19 and pregnancy is a new intercurrence in risk assessment for the care of the pregnant woman. Since the beginning of the pandemic in the country, cases have been increasing. The first case attended at the San Bartolomé Hospital was on April 12. Since the first reports of COVID+ pregnant women in China, to date, there is more important information on epidemiology and perinatal results for the management of the COVID+ pregnant woman. Objective: To determine the epidemiology and maternal perinatal outcomes of COVID-19 in pregnant women at the Hospital Nacional Docente Madre Niño San Bartolomé, Lima, Peru. Methods: Observational cross-sectional study, from April to July 2020. All the pregnant women who arrived at the obstetric emergency at Hospital San Bartolomé were included, and they underwent an immunochromatography test for IgM / IgG, to determine the seroprevalence of COVID-19. Obstetric and perinatal variables were collected in a data sheet upon admission to the emergency room. Results: Rapid test for SARS-CoV-19 was performed in 345 pregnant women who were hospitalized for delivery care. The average age was 27 years, with 10% adolescents and 16% over 35 years; 60% had 2 to 4 pregnancies, 38% of deliveries were vaginal, 15% of them preterm; 1.2% of the pregnant women were symptomatic and 0.2% were admitted to the intensive care unit. 61% of the newborns weighed between 2 500 and 3 500 g, 53% were male, 94% had an Apgar score greater than 7 at one minute, 3.3% with a positive swab within the first 24 hours. There were 3% fetal deaths. 48% of the pregnant women came from the Northern Cone of the city of Lima. Conclusions: Almost 100% of the pregnant women were asymptomatic and only 0.2% had respiratory complications. The mode of delivery by cesarean section was low in relation to other publications, all had obstetric indication. A small number of neonates had a positive swab. There was a higher incidence of fetal deaths in July 2020. The highest percentage of patients came from the Northern Cone of Lima.

Introduction: Community mitigation strategies (CMS) have demonstrated to be effective in the reduction of transmission and incidence of COVID-19, especially in the population with symptoms associated with the disease. The aim of this study was to evaluate the association between the presence of COVID-19 symptoms and adherence to CMS in Latin American adults. Methods: We carried out a secondary analysis of a database developed by the University of Maryland and Facebook social network during the COVID-19 pandemic. We included Latin American adults that used the Facebook platform and participated in a survey conducted from April 23 to May 23, 2020. The principal outcome variable was reported compliance with the three main CMS (physical distancing, use of face masks, and hand washing). The exposure variable included symptoms suspicious for COVID-19 defined as the presence of three or more symptoms of an acute clinical case of COVID-19. We performed generalized linear models of the Poisson family with a logarithmic link function to evaluate the association between the presence of COVID-19 symptoms and reported compliance with CMS. We calculated crude and adjusted prevalence ratios (PR) with their 95% confidence intervals (95%CI). Results: We analyzed 1,310,690 adults from Latin America; 48.1% were male and 42.9% were under 35 years of age. The prevalence of suspicious symptoms of COVID-19 was 18.5% and reported compliance with the three CMS was 45.3%. The countries with the highest proportion of reported compliance with the three CMS were Peru, Bolivia and Panama, while those with the lowest reported compliance were Costa Rica, Nicaragua and Honduras. In the adjusted model, people with suspicious symptoms for COVID-19 had a 14% lower compliance with the three CMS (aPR = 0.86; 95%CI: 0.85–0.87; p < 0.001). Conclusions: Less than half of the participants complied with the CMS, and those presenting suspicious symptoms for COVID-19 had lower reported compliance with the three CMS. / University of Maryland / Revisión por pares

El virus SARS-CoV-2 sigue reproduciéndose velozmente y muestra variantes más infecciosas. La segunda ola de la enfermedad va apaciguándose en Europa y Estados Unidos de América del Norte, menos en América del Sur. En el Perú, las cifras de fallecidos han sido sinceradas a más del doble, encontrándose mayor mortandad en los hombres y en los mayores de 70 años. Se ha corroborado en el mundo que la gestante sufre una enfermedad más severa -a veces con un proceso similar a la preeclampsia-, con más posibilidad de muerte que la mujer no gestante y mayor muerte fetal y prematuridad. Las vacunas iniciales elaboradas en los EE UU y Europa están siendo efectivas en disminuir las infecciones, hospitalizaciones y muertes en los países donde la vacunación ha avanzado más rápido. Está siendo administrada en gestantes sin efectos secundarios mayores, recomendándoseles se vacunen para evitar la infección severa. Mientras tanto, no se conoce la duración de la inmunidad dada por la infección COVID-19 y por la vacuna. Se está considerando una tercera dosis de refuerzo y el cambio de marca en los vacunados. Y se está en espera de una tercera ola de infecciones debido a la aparición de las variantes brasilera e india (delta). / The SARS-CoV-2 virus continues to reproduce rapidly and is showing more infectious variants. The second wave of the pandemic is subsiding in Europe and the United States of North America, but not in South America. In Peru, the number of deaths has more than doubled, with a higher mortality rate in men and in those over 70 years of age. It has been corroborated worldwide that pregnant women suffer a more severe disease -sometimes with a process similar to preeclampsia- with a greater possibility of death than non-pregnant women and greater fetal death and prematurity. Initial vaccines developed in the USA and Europe are proving effective in reducing infections, hospitalizations and deaths in countries where vaccination has advanced more rapidly. It is being administered in pregnant women without major side effects, and they are recommended to be vaccinated to avoid severe infection. As of now, the duration of immunity given by COVID-19 infection and by the vaccine is not known. A third booster dose and rebranding of vaccinees is being considered. And a third wave of infe ergence of the Brazilian and Indian (Delta) variants.

Introducción: El distrito de El Agustino pertenece a la DIRIS Lima Este con una población de 197,788 pobladores y un índice de pobreza de 22,10%. El principal problema de salud es el alto índice de morbilidad por IRAS y las neoplasias como la primera causa de mortalidad. Asimismo, el ascenso de casos de Covid-19 en el primer trimestre del año. Descripción del proyecto: Se planteó como objetivo mejorar la practica de estratégias preventivas frente al Covid-19 en la familia Meza Pucuhuayla. El total de participantes fueron 3 integrantes, un grupo heterogéneo en edades. Se elaboraron materiales educativos para todas las sesiones educativas y talleres, listas de cotejo para los monitoreos semanales. Se empleó la plataforma Kahoot para los pre y post test de las sesiones educativas. Resultados: Para las actividades planteadas se logró el 100% de eficacia y cobertura en las sesiones educativas con los temas de alimentación saludable, enfermedades respiratorias y generalidades del Covid-19. En relación a talleres todos tuvierton un volumen de 3 participantes instruidos y con una productividad en promedio de 10 minutos por cada integrante. Para la actividad de monitoreo de actividad fisica se logró un volumen de 6 listas de cotejo y una cobertura del 200%. Para el taller de bioseguridad y desinfección frente al Covid-19 se logró un volumen de 12 listas de cotejo monitoreadas. Conclusiones: Se logró incrementar prácticas de hábitos saludables para lo que se comparó en tres momentos las actividades físicas y elaboración de alimentación saludable. Asimismo, se logró identificar medidas preventivas para las enfermedades respiratorias prevalentes en sesiones educativas; se implementó un protocolo de desinfección y bioseguridad en el hogar en el cual se aprendió el correcto uso de desifecctantes, técnica de lavado de manos e inocuidad de alimentos. / Introduction: The district of El Agustino belongs to the East Lima DIRIS with a population of 197,788 inhabitants and a poorness rate of 22.10%. The main health problem is the high rate of morbidity due to ARIS and neoplasms as the first cause of mortality. Likewise, the rise in Covid-19 cases in the first quarter of the year. Description of project: The objective was to improve the practice of preventive strategies against Covid-19 in the Meza Pucuhuayla family. The total of participants were 3 members, a heterogeneous age group. Educational materials were prepared for all educational sessions and workshops, checklists for weekly monitoring. The Kahoot platform was used for the pre and post tests of the educational sessions. Results: For the proposed activities, 100% efficiency and coverage were achieved in the educational sessions with the topics of healthy eating, respiratory diseases and generalities of the Covid-19. In relation to workshops, all had a volume of 3 instructed participants and with an average productivity of 10 minutes for each member. For the physical activity monitoring activity, a volume of 6 checklists was achieved and a coverage of 200%. For the biosafety and disinfection workshop against Covid - 19, a volume of 12 monitored checklists was achieved. Conclusions: It was possible to increase the practices of healthy habits, for which physical activities and the preparation of healthy eating were compared in three moments. Likewise, it was possible to identify preventive measures for prevalent respiratory diseases in educational sessions; a disinfection and biosecurity protocol was implemented in the home in which the correct use of disinfectants, hand washing technique and food safety were learned. / Trabajo de investigación

Introduction: Only 3 types of coronavirus cause aggressive respiratory disease in humans (MERS-Cov, SARS-Cov-1, and SARS-Cov-2). It has been reported higher infection rates and severe manifestations (ICU admission, need for mechanical ventilation, and death) in patients with comorbidities such as diabetes mellitus (DM). For this reason, this study aimed to determine the prevalence of diabetes comorbidity and its associated unfavorable health outcomes in patients with acute respiratory syndromes for coronavirus disease according to virus types. Methods: Systematic review of literature in Pubmed/Medline, Scopus, Web of Science, Cochrane, and Scielo until April of 2020. We included cohort and cross-sectional studies with no restriction by language or geographical zone. The selection and extraction were undertaken by 2 reviewers, independently. The study quality was evaluated with Loney’s instrument and data were synthesized by random effects model meta-analysis. The heterogeneity was quantified using an I2 statistic. Funnel plot, Egger, and Begg tests were used to evaluate publication biases, and subgroups and sensitivity analyses were performed. Finally, we used the GRADE approach to assess the evidence certainty (PROSPERO: CRD42020178049). Results: We conducted the pooled analysis of 28 studies (n = 5960). The prevalence analysis according to virus type were 451.9 diabetes cases per 1000 infected patients (95% CI: 356.74-548.78; I2 = 89.71%) in MERS-Cov; 90.38 per 1000 (95% CI: 67.17-118.38) in SARS-Cov-1; and 100.42 per 1000 (95% CI: 77.85, 125.26 I2 = 67.94%) in SARS-Cov-2. The mortality rate were 36%, 6%, 10% and for MERS-Cov, SARS-Cov-1, and SARS-Cov-2, respectively. Due to the high risk of bias (75% of studies had very low quality), high heterogeneity (I2 higher than 60%), and publication bias (for MERS-Cov studies), we down rate the certainty to very low. Conclusion: The prevalence of DM in patients with acute respiratory syndrome due to coronaviruses is high, predominantly with MERS-Cov infection. The unfavorable health outcomes are frequent in this subset of patients. Well-powered and population-based studies are needed, including detailed DM clinical profile (such as glycemic control, DM complications, and treatment regimens), comorbidities, and SARS-Cov-2 evolution to reevaluate the worldwide prevalence of this comorbidity and to typify clinical phenotypes with differential risk within the subpopulation of DM patients. / Revisión por pares

Introducción: en la presente revisión conoceremos los detalles de esta nueva prueba de laboratorio, utilizada para cuantificar los anticuerpos neutralizantes contra el SARS-CoV-2. Esta prueba diagnóstica comienza a tener un mayor protagonismo, a razón del proceso de infección y vacunación en el mundo, para comprender los misterios del correlato de protección inmunológica. Contenido: los anticuerpos neutralizantes tienen la capacidad de bloquear la capacidad del virus, para unirse al receptor ACE2 en las células humanas y estos anticuerpos permiten eliminar el efecto de microorganismos invasores. Su actividad se genera por las proteínas situadas en la superficie de los virus, a las que se unen para «bloquear» la infección. Los anticuerpos neutralizantes se definen in vitro por su capacidad para bloquear la entrada, fusión o salida del coronavirus, es decir son anticuerpos funcionales. Resumen: en la actualidad existen diferentes pruebas de laboratorio (pruebas de inmunoensayo de alto rendimiento), que tienen la capacidad de detectar anticuerpos inmunoglobulinas G anti proteína S del SARS-CoV-2 y que se correlacionan con las pruebas de laboratorio gold standard para la determinación de estos anticuerpos. Es crucial que estas pruebas de inmunoensayo de alto rendimiento, sean validadas en su fabricación contra métodos gold standard para determinar la presencia de anticuerpos neutralizantes. Perspectiva: esta revisión pretende ampliar el conocimiento de esta nueva prueba, que en un futuro permitirán definir los valores de correlato inmunológico generados por las vacunas o por una infección previa.