Efficacy of some avian influenza H5 vaccines against local highly pathogenic avian influenza viruses subtype H5N8 isolated in 2018 and 2020 in Egypt

Commercial inactivated avian influenza H5 vaccine is used as an essential control strategy for avian influenza disease in Egypt. Since the initial outbreaks of highly pathogenic avian influenza H5N8, the virus has diverged with new genotypes and variant viruses continuing to emerge which mainly stand behind vaccination failure. In the present work, four different commercial avian influenza vaccines were inoculated in specific pathogenic free chickens for assessing its efficacy against local highly pathogenic avian influenza H5N8 virus isolated in 2018 and 2020. Two hundred and forty specific pathogenic free chickens were clustered into four groups; each group was inoculated with the corresponding vaccine (60 specific pathogenic free chickens/vaccine). Sixty specific pathogenic free chicks were kept as control unvaccinated group. Sera collected from vaccinated chicken groups at 3rd and 4th week post vaccination were examined for calculating neutralizing antibodies using heterologous highly pathogenic avian influenza H5N8 2018 and 2020. At 4th week post vaccination, vaccinated chickens were challenged; moreover, oropharyngeal swabs were collected from challenged vaccinated chickens to calculate the viral shedding. Our findings revealed the groups vaccinated with vaccine code no 1 and 2 that contains two vaccine strains (H5N1 and H5N8) of local origin exhibited the highest hemagglutination inhibition titer, protection (percent) and reduction in viral shedding titer when examined by highly pathogenic avian influenza H5N8 2018 while, vaccine code no 3 induced lower antibody response, protection (percent) and reduction in viral shedding, but still within satisfactory level when compared to previous groups. When highly pathogenic avian influenza H5N8 2020 was used, it was found the seroconversion rate, protection (percent) and mean titer of reduction of viral shedding decreased in comparison to those recorded for highly pathogenic avian influenza H5N8 2018. Vaccine code no 4 was impotent to either highly pathogenic avian influenza 2018 or 2020. Accordingly, it was recommended to update vaccine strain according to epidemiological condition and used the predominant circulating strain isolate in challenge test(AU)

La vacuna comercial inactivada H5 se utiliza como estrategia esencial de control de la enfermedad de la gripe aviar en Egipto. Desde los brotes iniciales de la gripe aviar altamente patógena H5N8, el virus ha variado al aparecer continuamente nuevos genotipos y variantes virales, que son los principales responsables del fracaso de la vacunación. En el presente trabajo, cuatro vacunas comerciales diferentes contra la gripe aviar se inocularon en pollos libres de patógenos específicos para evaluar su eficacia contra cepas del virus local de la gripe aviar altamente patógeno H5N8 aisladas en 2018 y 2020. Se agruparon 240 pollos pollos libres de patógenos específicos en cuatro grupos, cada uno fue inoculado con la vacuna correspondiente (60 pollos pollos libres de patógenos específicos/vacuna). Sesenta pollos SPF se mantuvieron como grupo control sin vacunar. Los sueros de los pollos vacunados recogidos en la 3ª y 4ª semana después de la vacunación se examinaron para calcular los anticuerpos neutralizantes contra la gripe aviar heteróloga H5N8 2018 y 2020. En la cuarta semana después de la vacunación, los pollos vacunados fueron retados; además, se recogieron hisopados orofaríngeos de los pollos vacunados retados para calcular la diseminación viral. Nuestros resultados revelaron que los grupos vacunados con las vacunas con códigos nº 1 y 2, que contienen dos cepas vacunales (H5N1 y H5N8) de origen local, mostraron el mayor título de inhibición de la hemaglutinación, protección (por ciento) y reducción del título de excreción viral cuando se evaluaron contra la gripe aviar altamente patógena H5N8 2018, mientras que la vacuna con código nº 3 indujo menor respuesta de anticuerpos, protección (por ciento) y reducción de la excreción viral, pero todavía dentro de un nivel satisfactorio en comparación con los grupos anteriores. Al utilizar la vacuna contra la gripe aviar altamente patógena H5N8 2020, se observó que la tasa de seronconversión, la protección (por ciento) y el título medio de reducción de la excreción viral disminuyeron en comparación con los registrados para la gripe aviar altamente patógena H5N8 2018. La vacuna con código nº 4 no fue potente para la gripe aviar altamente patógena de 2018 o de 2020. Por consiguiente, se recomendó actualizar la cepa de la vacuna de acuerdo con las condiciones epidemiológicas y utilizar el aislamiento de la cepa circulante predominante en la prueba de reto(AU)

Métodos diagnósticos para la infección por SARS-CoV-2 / Diagnostic methods for SARS-CoV-2 infection

En diciembre de 2019 se identificó el virus SARS-CoV-2, cuya rápida propagación global puso en estado de emergencia al mundo entero, llevando al ser humano a una situación sin antecedente cercano. El objetivo de esta revisión es describir los métodos diagnósticos utilizados actualmente para identificar la infección por SARS-CoV-2. Las manifestaciones clínicas y el espectro imagenológico de la enfermedad son muy inespecíficos y no permiten realizar un diagnóstico certero. Por esta razón, es esencial una apropiada toma de muestra respiratoria en el momento y sitio anatómico adecuado para un diagnóstico preciso de COVID-19. La técnica de muestreo más utilizada es el hisopado nasofaríngeo y la prueba diagnóstica más fiable se basa en la retrotranscripción seguida por reacción en cadena de la polimerasa en tiempo real (RT-PCR). No obstante, existen otras técnicas moleculares, como también tests serológicos para detectar anticuerpos o fragmentos antigénicos del SARS-CoV-2. Más allá de la precisión diagnóstica, es importante tener en cuenta la probabilidad basal (pretest) para interpretar correctamente el resultado obtenido y aislar aquellos posibles falsos negativos. Con el objetivo de evitar la saturación del sistema de salud es imprescindible contar con información y métodos diagnósticos precisos para detectar tempranamente los focos de infección y reducir la transmisión comunitaria, utilizando eficazmente los diferentes recursos diagnósticos. (AU)

In December 2019, the SARS-CoV-2 virus was identified for the first time, whose rapid global spread put the entire world in a state of emergency, leading humans to an unprecedented situation with no immediate history. The main purpose of this review is to describe the diagnostic methods currently used to identify SARS-CoV-2 infection. The clinical manifestations and the imaging spectrum of the disease are nonspecific and do not allow an accurate diagnosis to be made. For this reason, an appropriate respiratory sampling at the right time and anatomical site is essential for an accurate diagnosis of COVID-19. The most widely used sampling technique is nasopharyngeal swab, and the most reliable diagnostic test is by reverse transcription followed by real-time polymerase chain reaction (RT-PCR). However, there are other molecular techniques, as well as serological tests to detect antibodies or antigenic fragments of SARS-CoV-2. Beyond the diagnostic precision, it is important to take into account the baseline probability (pre-test) to correctly interpret the result obtained and isolate those possible false negatives. In order to avoid saturation of the health system, it is essential to have accurate information and diagnostic methods to detect outbreaks of infection in early stages and to reduce communitary transmission, making effective use of the various diagnostic resources. Coronavirus infections/diagnosis, viral/diagnosis, pandemics, clinical laboratory techniques, real-time polymerase chain reaction, antigens, viral/analysis. (AU)

Evaluación en condiciones de campo de una prueba serológica rápida para detección de anticuerpos IgM e IgG contra SARS-CoV-2 / Evaluation of a rapid serological test for detection of IgM and igG antibodies against SARS-CoV-2 under field conditions

RESUMEN Objetivos: Determinar el rendimiento diagnóstico adicional de una prueba serológica rápida que detecta anticuerpos IgM e IgG contra SARS-CoV-2 en relación a la reacción en cadena de polimerasa reversa en tiempo real (RT-PCR). Materiales y métodos: Se realizó un estudio transversal incluyendo pacientes hospitalizados por COVID-19 en tres hospitales, trabajadores de salud expuestos a la infección y pacientes ambulatorios que cumplían criterios de caso sospechoso, a quienes se les realizó la prueba molecular (RT-PCR) y la prueba serológica rápida. Se evaluó el rendimiento diagnóstico adicional de las prueba serológica rápida en relación a la molecular. Asimismo, se realizó la estimación de sensibilidad y especificidad de dichas pruebas. Resultados: Se incluyeron 144 personas. La prueba serológica rápida obtuvo un 19,4% de resultados positivos en comparación con un 11,1% en la prueba molecular (p=0,03). La prueba serológica rápida detectó 21 casos que habían resultado negativos por el RT-PCR inicial y el rendimiento diagnóstico adicional fue de 56,8% en comparación al RT-PCR. El rendimiento diagnóstico adicional fue 50,0% durante la primera semana, 70,0% durante la segunda y 50,0% durante la tercera semana de inicio de síntomas. La sensibilidad de la prueba serológica rápida fue de 43,8% y la especificidad del 98,9%. Conclusiones: La prueba serológica rápida logró detectar un mayor número de casos respecto a la molecular, sobre todo a partir de la segunda semana de inicio de síntomas. Además, presentó una alta especificidad. Los resultados mostrarían su utilidad como prueba complementaria a la prueba molecular, especialmente durante la segunda y tercera semana de enfermedad.

ABSTRACT Objective: To determine the additional diagnostic performance of a rapid serological test for detection of IgM and IgG antibodies compared to the real-time polymerase chain reaction (RT-PCR) test; for detection of SARS-CoV-2. Materials and methods: A cross-sectional study was carried out including patients hospitalized for COVID-19 in 3 hospitals, health workers exposed to the infection and outpatients who met suspicious case criteria, all of which underwent the molecular test (RT-PCR) and the rapid serological test. The additional diagnostic performance of rapid serological test was evaluated in comparison to molecular tests. Likewise, an approximation was made to the sensitivity and specificity of the rapid serological test. Results: 144 people were included. With the rapid test, 19.4% of positive results were obtained compared to 11.1% in the molecular test (p = 0.03). The rapid serological test detected 21 cases that had been negative by the initial (RT-PCR), providing an additional diagnostic performance of 56.8% compared to the RT-PCR. The additional diagnostic performance was 50.0% during the first week, 70.0% during the second week and 50.0% during the third week of symptom onset. The sensitivity of the rapid serological test was 43.8% and the specificity of 98.9%. Conclusions: The rapid serological test was able to detect a greater number of cases than those detected by the molecular test especially after the second week of onset of symptoms. It also showed high specificity. It is therefore useful as a complementary test to RT-PCR, especially during the second and third week of illness.

Diagnóstico microbiológico de SARS-COV 2 / Microbiological diagnosis of SARS-VOC 2

Introducción: La enfermedad producida por el nuevo coronavirus constituye un reto para los sistemas de salud. En estos tiempos de pandemia disponer de pruebas que ayuden a un diagnóstico temprano e incluso a detectar pacientes asintomáticos, es una de las claves para disminuir los contagios y evitar la propagación. Objetivo: Revisar los aspectos más importantes en el diagnóstico del nuevo coronavirus. Desarrollo: La detección de ARN de SARS-CoV2 en muestras respiratorias, es la técnica de referencia y de elección para el diagnóstico microbiológico de COVID-19. Tomando la muestra de la parte posterior de la faringe y de las fosas nasales puede detectarse la presencia del virus. La detección de antígenos es un tipo de prueba de diagnóstico rápido la cual detecta la presencia de proteínas virales (antígenos) expresadas por el virus de la COVID-19. La detección de los anticuerpos generados en el organismo huésped infectado es una de las técnicas más utilizadas a nivel mundial en grandes poblaciones, incluso como pesquizaje, aunque su interpretación puede requerir intervención de médicos especializados. También está basada en la detección de anticuerpos del tipo IgM e IgG y algunas presentan la detección de anticuerpos IgA. Conclusiones: La interpretación de las pruebas serológicas debe realizarse con cautela, teniendo en cuenta sus limitaciones, y evaluarlas acorde a la situación clínica del paciente y de los resultados de la prueba de referencia(AU)

Introduction: The disease caused by the new coronavirus constitutes a challenge for health systems. In these times of pandemic, having tests that help early diagnosis and even detect asymptomatic patients is one of the keys to reducing infections and preventing the spread. Objective: To review the most important aspects in the diagnosis of the new coronavirus. Findings: Detection of SARS-CoV2 RNA in respiratory samples is the reference and technique of choice for the microbiological diagnosis of COVID-19. By taking samples from the back of the pharynx and the nostrils, the presence of the virus can be detected. Antigen detection is a type of rapid diagnostic test which detects the presence of viral proteins (antigens) expressed by COVID-19 virus. Detection of the antibodies generated in the infected host organism is one of the most widely used techniques worldwide in large populations, even as screening, although its interpretation may require the intervention of specialists. It is also based on the detection of IgM and IgG type antibodies and some have the detection of IgA antibodies. Conclusions: The interpretation of serological tests should be done with caution, taking into account the limitations, and assessing them according to the patient's clinical situation and the results of the reference test(AU)

Performance of the Dot-blot test method for detecting antibodies to Sarcocystis spp. in cattle / Desempenho do teste Dot-blot para detecção de anticorpos para Sarcocystis spp. em bovinos

Serological techniques can detect antibodies against Sarcocystis spp., Neospora caninum and Toxoplasma gondii antigens in single or mixed infections. Immunofluorescent antibody tests (IFAT) is considered the gold standard technique for Sarcocystosis diagnostic in cattle serum and a positive IFAT result reflects Sarcocystis spp. infection. Therefore, the aims of the present study were to compare IFAT and Dot-blot for sarcocystosis diagnostic in experimentally infected mice and to investigate serological cross-reactions with N. caninum and T. gondii in these methods. Mice (Mus musculus) were inoculated intraperitoneally with bradizoites of Sarcocystis spp. or tachyzoites of N. caninum or T. gondii. Serum samples were obtained and analyzed by IFAT and Dot-blot for the three protozoa. Serum from N. caninum and T. gondii experimentally infected mice were tested by IFAT and reacted only to N. caninum or T. gondii antigens, respectively. Specific antibodies against Sarcocystis spp. were present in all animals experimentally infected with this protozoan, with IFAT titers from 10 to 800. Serum samples from mice experimentally infected with Sarcocystis spp., N. caninum and T. gondii and tested by Dot-blot demonstrated no cross reaction between protozoa. A Dot-blot using Sarcocystis spp. antigen appears to be a good alternative to IFAT in the serological diagnosis of Sarcocystosis.(AU)

As técnicas sorológicas podem detectar anticorpos contra os antígenos de Sarcocystis spp., Neospora caninum e Toxoplasma gondii em infecções únicas ou mistas. O teste de anticorpos imunofluorescentes (IFAT) é considerado a técnica padrão-ouro para o diagnóstico de sarcocistose no soro de bovinos e um resultado positivo de IFAT reflete Sarcocystis spp. infecção. Portanto, os objetivos do presente estudo foram comparar IFAT e Dot-blot para diagnóstico de sarcocistose em camundongos infectados experimentalmente e investigar reações cruzadas sorológicas com N. caninum e T. gondii nesses métodos. Os camundongos (Mus musculus) foram inoculados intraperitonealmente com bradizoítos de Sarcocystis spp. ou taquizoítos de N. caninum ou T. gondii. As amostras de soro foram obtidas e analisadas por IFAT e Dot-blot para os três protozoários. O soro de N. caninum e T. gondii infectados experimentalmente foram testados por IFAT e reagiram apenas aos antígenos de N. caninum ou T. gondii, respectivamente. Anticorpos específicos contra Sarcocystis spp. estavam presentes em todos os animais experimentalmente infectados com este protozoário, com títulos de IFAT de 10 a 800. Amostras de soro de camundongos infectados experimentalmente com Sarcocystis spp., N. caninum e T. gondii e testadas por Dot-blot não demonstraram reação cruzada entre protozoários. Um Dot-blot usando Sarcocystis spp. O antígeno parece ser uma boa alternativa ao IFAT no diagnóstico sorológico da sarcocistose.(AU)

Plano municipal de contingência de Araguacema-Tocantins: Novo Coronavírus (covid-19) - segunda versão / Municipal contingency plan of Araguacema-Tocantins: New Coronavirus (covid-19) - second version

Orienta na campanha para ações de combate ao Coronavírus (Covid-19) no município de Araguacema no Tocantins. Apresenta quais as definições de casos de infecção humana pelo COVID-19. Orientações de como notificar ao Centro de Informações Estratégicas de Vigilância em Saúde (CIEVS). Quais os períodos de incubação da doença. Fatores sobre a transmissão e tratamento. Investigação epidemiológica. Quais atribuições da Vigilância em Saúde. Orientações para a coleta de amostras no Laboratório Central de Saúde Pública do Tocantins (LACEN-TO) bem como a técnica de coleta de Swabde nasofaringe e orofaringe (swabs combinados), o acondicionamento, transporte e envio das amostras. Traz as recomendações para a coleta de amostras em situação de óbito. Mostra as medidas de prevenção e controle Precauções padrão, as medidas de isolamento. Transporte do paciente. Como se dá a Limpeza e desinfecção de superfícies. Quais as atribuições das unidades hospitalares quanto aos cuidados com o paciente.

He guides in the campaign for actions to combat the Coronavirus (Covid-19) in the municipality of Araguacema in Tocantins. It presents the definitions of cases of human infection by COVID-19. Guidelines on how to notify the Health Surveillance Strategic Information Center (CIEVS). What are the disease incubation periods. Factors about transmission and treatment. Epidemiological investigation. Which attributions of Health Surveillance. Guidelines for the collection of samples at the Central Laboratory of Public Health of Tocantins (LACEN-TO) as well as the technique of collecting Swabde nasopharynx and oropharynx (combined swabs), packaging, transport and sending of samples . It provides recommendations for the collection of samples in situations of death. Shows prevention and control measures Standard precautions, isolation measures. Transporting the patient. How to clean and disinfect surfaces. What are the attributions of hospital units regarding patient care.

Orienta en la campaña de acciones de combate al Coronavirus (Covid-19) en el municipio de Araguacema en Tocantins. Presenta las definiciones de casos de infección humana por COVID-19. Directrices sobre cómo notificar al Centro de Información Estratégica de Vigilancia Sanitaria (CIEVS). Cuáles son los períodos de incubación de la enfermedad. Factores de transmisión y tratamiento. Investigación epidemiológica. Qué atribuciones de la Vigilancia Sanitaria. Lineamientos para la recolección de muestras en el Laboratorio Central de Salud Pública de Tocantins (LACEN-TO) así como la técnica de recolección de Swabde nasofaringe y orofaringe (hisopos combinados), el empaque, transporte y envío de las muestras . Proporciona recomendaciones para la recolección de muestras en situaciones de muerte. Muestra medidas de prevención y control Precauciones estándar, medidas de aislamiento. Transporte del paciente. Cómo limpiar y desinfectar superficies. Cuáles son las atribuciones de las unidades hospitalarias en cuanto a la atención al paciente.

Il guide dans la campagne d'actions de lutte contre le Coronavirus (Covid-19) dans la commune d'Araguacema à Tocantins. Il présente les définitions des cas d'infection humaine par COVID-19. Lignes directrices sur la manière de notifier le Centre d'information stratégique de surveillance sanitaire (CIEVS). Quelles sont les périodes d'incubation de la maladie. Facteurs de transmission et de traitement. Enquête épidémiologique. Quelles attributions de la Surveillance de la Santé. Directives pour le prélèvement d'échantillons au Laboratoire Central de Santé Publique de Tocantins (LACEN-TO) ainsi que la technique de prélèvement de Swabde nasopharynx et oropharynx (écouvillons combinés), l'emballage, le transport et l'envoi des échantillons . Il fournit des recommandations pour le prélèvement d'échantillons en cas de décès. Affiche les mesures de prévention et de contrôle Précautions standard, mesures d'isolement. Transport du patient. Comment nettoyer et désinfecter les surfaces. Quelles sont les attributions des unités hospitalières en matière de soins aux patients.

The direct and indirect effects of the pneumococcal conjugated vaccine on carriage rates in children aged younger than 5 years in Latin America and the Caribbean: a systematic review / Efeitos diretos e indiretos da vacina pneumocócica conjugada no estado de portador em crianças menores de 5 anos, na América Latina e no Caribe: uma revisão sistemática

ABSTRACT Objective To demonstrate the impact of pneumococcal conjugate vaccine in Streptococcus pneumoniae carriage status in children younger than 5 years in Latin America and the Caribbean. Methods A systematic literature review was carried out on the direct and indirect effects of pneumococcal vaccine in the carriage status, after implementation in childhood immunization programs. Studies carried out in children younger than 5 years were selected from the PubMed® and Virtual Health Library databases, and data collected after implementation of pneumococcal vaccine in Latin America and the Caribbean, between 2008 and 2018. Results From 1,396 articles identified, 738 were selected based on titles and abstracts. After duplicate removal, 31 studies were eligible for full-text reading, resulting in 6 publications for analysis. All selected publications were observational studies and indicated a decrease in the carriage and vaccine types, and an increase in the circulation of non-vaccine serotypes, such as 6A, 19A, 35B, 21 and 38. We did not identify changes in the antimicrobial resistance after vaccine implementation. Conclusion A decrease in the carriage status of vaccine types and non-vaccine types was detected. The continuous monitoring of pneumococcal vaccine effect is fundamental to demonstrate the impact of the carriage status and, consequently, of invasive pneumococcal disease, allowing better targeting approaches in countries that included pneumococcal vaccine in their immunization programs. Our study protocol was registered in PROSPERO (www.crd.york.ac.uk/prospero) under number CRD42018096719.

RESUMO Objetivo Demonstrar o impacto das vacinas pneumocócicas conjugadas no estado de portador de Streptococcus pneumoniae em crianças menores de 5 anos na América Latina e no Caribe. Métodos Foi realizada revisão sistemática da literatura sobre os efeitos diretos e indiretos da vacina pneumocócica no estado de portador em crianças menores de 5 anos, após a implantação da vacina nos calendários de imunização infantil. A partir de dados da PubMed®e da Biblioteca Virtual da Saúde, foram selecionados estudos de portador em crianças menores de 5 anos, com dados coletados após implementação da vacina de 2008 a 2018, na América Latina e no Caribe. Resultados Dos 1.396 artigos identificados, 738 foram selecionados mediante leitura de títulos e resumos. Após a extração dos duplicados, 31 foram elegíveis para leitura do texto completo, restando 6 artigos para análise. Todos os estudos selecionados eram observacionais e indicavam diminuição do portador e tipos vacinais, e aumento da circulação de sorotipos não vacinais, como 6A, 19A, 35B, 21 e 38. Não foi observada alteração na resistência antimicrobiana após a introdução da vacina. Conclusão Detectou-se redução no estado de portador, dos tipos vacinais e não vacinais. O monitoramento contínuo do efeito das vacinas pneumocócicas é fundamental, para demonstrar o impacto do estado de portador e, consequentemente, da doença pneumocócica invasiva, permitindo o melhor direcionamento nas ações em saúde para os países que incluíram a vacina no calendário de imunização. Nosso protocolo de estudo foi registrado no PROSPERO (www.crd.york.ac.uk/prospero) sob o número CRD42018096719.

Performance evaluation of an indirect immunofluorescence kit for the serological diagnosis of dengue / Avaliação do desempenho de kit de imunofluorescência indireta para o diagnóstico sorológico de dengue

ABSTRACT Objective: To evaluate the performance of indirect immunofluorescence for serological diagnosis of dengue virus in a population with high prevalence of arboviruses. Methods: Two-hundred serum samples from patients with clinical suspicion of dengue fever were tested by immunoenzymatic and indirect immunofluorescence assay BIOCHIP® mosaic. Specificity, sensitivity and Kappa coefficient were calculated. Discordant samples were tested by polymerase chain reaction for confirmation. Results: Of the 200 samples, 20% were positive and 80% negative for anti-dengue virus IgM antibodies in the immunoenzymatic test. Of the 40 positives, 25% were negative in indirect immunofluorescence. Of these ten discordant results, only 20% were also negative in the polymerase chain reaction (PCR). Of the 160 negatives in the immunoenzymatic test, 5% were positive in indirect immunofluorescence. Of these nine discordant results, 33% were positive in the PCR. The Kappa coefficient was 0.7 (0.572-0.829). Sensitivity and specificity of indirect immunofluorescence were respectively 75% and 94%. For anti-dengue virus IgG antibodies, of the 200 samples, 15.5% were positive and 84.5% were negative in the immunoenzymatic test. Of the 31 positives, 12.9% were negative in indirect immunofluorescence. Of these four discordant results, 25% were negative in the PCR. Of the 169 negatives, 8% were positive in indirect immunofluorescence. Of these 14 discordant results, 64% were also positive in the PCR. The Kappa coefficient was 0.695 (0.563-0.83). Sensitivity and specificity of indirect immunofluorescence were 87.1% and 91.7%, respectively. Conclusion: For diagnosis of acute infection, the immunoenzymatic test is enough, and the use of additional methods is not warranted. Replacing the immunoenzymatic test by indirect immunofluorescence would compromise the sensitivity for IgM. However, indirect immunofluorescence can distinguish three arboviruses simultaneously, an advantage during concomitant epidemics.

RESUMO Objetivo: Avaliar o desempenho da imunofluorescência indireta no diagnóstico sorológico de dengue em uma população com alta prevalência de arboviroses. Métodos: Duzentas amostras de soro de pacientes com suspeita clínica de dengue foram testadas por ensaio imunoenzimático e imunofluorescência indireta mosaico BIOCHIP®. Foram calculados especificidade, sensibilidade e coeficiente Kappa. Nas amostras discordantes, realizou-se reação em cadeia da polimerase como método confirmatório. Resultados: Das 200 amostras, 20% foram positivas e 80% negativas para IgM antivírus da dengue no ensaio imunoenzimático. Das 40 positivas, 25% foram negativas na imunofluorescência indireta. Destas dez negativas, apenas 20% eram também negativas na reação em cadeia da polimerase. Das 160 negativas no ensaio imunoenzimático, 5% foram positivas na imunofluorescência indireta. Por fim, dentre as nove discordantes, 33% tiveram vírus da dengue detectado na reação em cadeia da polimerase. O coeficiente Kappa foi 0,70 (0,57-0,82). Sensibilidade e especificidade por imunofluorescência indireta foram, respectivamente, 75% e 94%. Para IgG antivírus da dengue, de 200 amostras, 15,5% foram positivas e 84,5% negativas no ensaio imunoenzimático. Das 31 positivas, 12,9% foram negativas na imunofluorescência indireta. Destas quatro discordantes, 25% apresentaram vírus da dengue não detectado na reação em cadeia da polimerase. Das 169 negativas, 8% foram positivas na imunofluorescência indireta. Destas, 64% foram positivas também na reação em cadeia da polimerase. O coeficiente Kappa foi 0,695 (0,56-0,83). Sensibilidade e a especificidade por imunofluorescência indireta foram, respectivamente, 87,1% e 91,7%. Conclusão: Ensaio imunoenzimático seria suficiente para diagnóstico sorológico de infecção aguda, não justificando a incorporação da imunofluorescência indireta. Substituir ensaio imunoenzimático pela imunofluorescência indireta poderia comprometer a sensibilidade para IgM. Contudo, a imunofluorescência indireta auxilia diferenciar três arboviroses simultaneamente, sendo vantajoso em epidemias concomitantes.

Zika virus serological diagnosis: commercial tests and monoclonal antibodies as tools

Zika virus (ZIKV), an emerging arthropod-borne virus (arbovirus) of the Flaviviridae family, is a current issue worldwide, particularly because of the congenital and neurological syndromes associated with infection by this virus. As the initial clinical symptoms of all diseases caused by this group are very similar, clinical diagnosis is difficult. Furthermore, laboratory diagnostic efforts have failed to identify specific and accurate tests for each virus of the Flaviviridae family due to the cross-reactivity of these viruses in serum samples. This situation has resulted in underreporting of the diseases caused by flaviviruses. However, many companies developed commercial diagnostic tests after the recent ZIKV outbreak. Moreover, health regulatory agencies have approved different commercial tests to extend the monitoring of ZIKV infections. Considering that a specific and sensitive diagnostic method for estimating risk and evaluating ZIKV propagation is still needed, this review aims to provide an update of the main commercially approved serological diagnostics test by the US Food and Drug Administration (FDA) and Brazilian National Health Surveillance Agency (ANVISA). Additionally, we present the technologies used for monoclonal antibody production as a tool for the development of diagnostic tests and applications of these antibodies in detecting ZIKV infections worldwide.(AU)

Application of WHO International Biological Reference Standards to evaluate commercial serological tests for chronic Chagas disease

BACKGROUND Chagas disease, resulting from Trypanosoma cruzi infections, continues to be a health concern mainly in Latin American countries where the parasite is endemic. The laboratory diagnosis of a chronic infection is determined through serological assays for antibodies against T. cruzi and several tests are available that differ in key components, formats and methodologies. To date, no single test meets the criteria of a gold standard. The situation is further complicated by the difficulties associated with performance comparisons between different immunoassays or methodologies executed at different times and geographical areas. OBJECTIVE To improve the diagnosis of Chagas disease, the WHO coordinated the development of two International Biological Reference Standards for antibodies against anti-T. cruzi: NIBSC 09/186 and NIBSC 09/188 that respectively represent geographical regions with the highest prevalence of TcII and TcI lineages of the parasite. METHODS The principle goal of this study was to verify the behavior of these standards when assayed by several commercially available serological tests that employ different methods to capture and detect human anti-T. cruzi antibodies. FINDINGS AND MAIN CONCLUSIONS The results reinforce the recommendation that these standards be considered for performance evaluations of commercialised immunoassays and should be an integral step in the development of new test components or assay paradigms.

Plano de Contingência Municipal para Enfrentamento pelo Novo Coronavírus COVID-19 / Municipal Contingency Plan for Coping with the New Coronavirus COVID-19

Orienta na campanha para ações de combate ao Coronavírus (Covid-19) no município de Araguaína no Tocantins. Defini responsabilidades e prioridades na esfera municipal e local, assim como também ajuda organizar o fluxograma de resposta às emergências em saúde pública. Defini fluxos de referência para atendimento aos casos suspeitos com sintomas respiratórios leves, moderados e graves. Traz parâmetros para ajudar a detectar, identificar e notificar todos os casos suspeitos de coronavírus (COVID-19). Orienta no fluxo de vigilância epidemiológica para o diagnóstico dos casos suspeitos. Busca orientar e recomendar medidas de controle e prevenção da doença, de forma ativa, imediata e oportuna. Tenta estabelecer cuidados para redução do risco geral de contaminação pelo COVID-19 aos profissionais envolvidos nos atendimentos e protocolos relacionados. Promove ações de educação em saúde e orienta na divulgação das informações.

He guides in the campaign for actions to combat the Coronavirus (Covid-19) in the municipality of Araguaína in Tocantins. I defined responsibilities and priorities at the municipal and local level, as well as helping to organize the flowchart for responding to public health emergencies. I defined reference flows to care for suspected cases with mild, moderate and severe respiratory symptoms. It provides parameters to help detect, identify and report all suspected cases of coronavirus (COVID-19). Guides in the flow of epidemiological surveillance for the diagnosis of suspected cases. It seeks to guide and recommend disease control and prevention measures, in an active, immediate and timely manner. Attempts to establish care to reduce the general risk of contamination by COVID-19 to the professionals involved in the services and related protocols. It promotes health education actions and guides the dissemination of information.

Orienta en la campaña de acciones de combate al Coronavirus (Covid-19) en el municipio de Araguaína en Tocantins. Definí responsabilidades y prioridades a nivel municipal y local, además de ayudar a organizar el diagrama de flujo para responder a emergencias de salud pública. Definí flujos de referencia para la atención de casos sospechosos con sintomatología respiratoria leve, moderada y grave. Proporciona parámetros para ayudar a detectar, identificar y notificar todos los casos sospechosos de coronavirus (COVID-19). Guías en el flujo de vigilancia epidemiológica para el diagnóstico de casos sospechosos. Busca orientar y recomendar medidas de control y prevención de enfermedades, de manera activa, inmediata y oportuna. Intentos de establecer cuidados para reducir el riesgo general de contaminación por COVID-19 a los profesionales involucrados en los servicios y protocolos relacionados. Promueve acciones de educación para la salud y orienta la difusión de información.

Il guide dans la campagne d'actions de lutte contre le Coronavirus (Covid-19) dans la municipalité d'Araguaína à Tocantins. J'ai défini les responsabilités et les priorités aux niveaux municipal et local, ainsi que d'aider à organiser l'organigramme pour répondre aux urgences de santé publique. J'ai défini des flux de référence pour prendre en charge les cas suspects présentant des symptômes respiratoires légers, modérés et sévères. Il fournit des paramètres pour aider à détecter, identifier et signaler tous les cas suspects de coronavirus (COVID-19). Guides dans le flux de la surveillance épidémiologique pour le diagnostic des cas suspects. Il cherche à guider et à recommander des mesures de contrôle et de prévention des maladies, de manière active, immédiate et opportune. Tentative de mise en place de soins pour réduire le risque général de contamination par COVID-19 pour les professionnels impliqués dans les services et les protocoles associés. Il promeut les actions d'éducation sanitaire et guide la diffusion de l'information.

Plano de Contingência Municipal para Enfrentamento ao Novo Coronavírus COVID-19 / Municipal contingency plan to Confront the new Coronavirus COVID-19

Orienta na campanha para ações de combate ao Coronavírus (Covid-19) no município de Araguatins no Tocantins. Orienta o Sistema de Vigilância em Saúde e a Rede de Unidades de Saúde do município para atuação na identificação, notificação e investigação de casos suspeitos de Doença Respiratória Aguda pelo 2019-nCoV, de modo a evitar os riscos de transmissão. Descreve estratégias de identificação oportuna de casos suspeitos, conforme preconizado pelo Ministério da Saúde (MS), no sentido de controlar e reduzir a disseminação do Covid-19 no município. Definir responsabilidades e prioridades na esfera municipal, assim como também organizar o fluxograma de resposta às emergências em saúde pública. Orienta e recomenda medidas de controle e prevenção da doença, de forma ativa, imediata e oportuna. Defini fluxos de referência para atendimento aos casos suspeitos com sintomas respiratórios leves e graves

He guides in the campaign for actions to combat the Coronavirus (Covid-19) in the municipality of Araguatins in Tocantins. It guides the Health Surveillance System and the Network of Health Units in the municipality to act in the identification, notification and investigation of suspected cases of Acute Respiratory Disease by 2019-nCoV, in order to avoid the risks of transmission. It describes strategies for the timely identification of suspected cases, as recommended by the Ministry of Health (MS), in order to control and reduce the spread of Covid-19 in the municipality. Define responsibilities and priorities at the municipal level, as well as organize the flowchart for responding to public health emergencies. It guides and recommends measures to control and prevent the disease, in an active, immediate and timely manner. Defined reference flows for care of suspected cases with mild and severe respiratory symptoms

Orienta en la campaña de acciones de combate al Coronavirus (Covid-19) en el municipio de Araguatins en Tocantins. Orienta al Sistema de Vigilancia en Salud y a la Red de Unidades de Salud del municipio para actuar en la identificación, notificación e investigación de casos sospechosos de Enfermedad Respiratoria Aguda para el 2019-nCoV, con el fin de evitar los riesgos de transmisión. Describe estrategias para la identificación oportuna de casos sospechosos, según lo recomendado por el Ministerio de Salud (MS), con el fin de controlar y reducir la propagación de Covid-19 en el municipio. Definir responsabilidades y prioridades a nivel municipal, así como organizar el diagrama de flujo para responder a emergencias de salud pública. Orienta y recomienda medidas para el control y prevención de la enfermedad, de forma activa, inmediata y oportuna. Flujos de referencia definidos para la atención de casos sospechosos con síntomas respiratorios leves y graves

Il guide dans la campagne d'actions de lutte contre le Coronavirus (Covid-19) dans la commune d'Araguatins à Tocantins. Il guide le système de surveillance sanitaire et le réseau des unités de santé de la commune pour agir dans l'identification, la notification et l'enquête des cas suspects de maladie respiratoire aiguë d'ici 2019-nCoV, afin d'éviter les risques de transmission. Il décrit les stratégies d'identification rapide des cas suspects, comme le recommande le ministère de la Santé (MS), afin de contrôler et de réduire la propagation du Covid-19 dans la municipalité. Définir les responsabilités et les priorités au niveau municipal, et organiser l'organigramme pour répondre aux urgences de santé publique. Il oriente et recommande des mesures pour contrôler et prévenir la maladie, de manière active, immédiate et opportune. Flux de référence définis pour la prise en charge des cas suspects présentant des symptômes respiratoires légers et graves

Control de calidad en pruebas cualitativas aplicado a pruebas sexológicas / Quality control in qualitative tests applied to sexological tests

Si bien el control de calidad en determinaciones cuantitativas tiene amplia difusión, hay extensa bibliografía que lo avala y está en constante actualización, no sucede lo mismo cuando se refiere a pruebas cualitativas como las que se utilizan en diagnósticos serológicos. Es importante antes de la utilización de esta prueba serológica en el laboratorio disponer de información acerca de su caracterización, utilidad y algoritmos de trabajo más convenientes. Si bien se cuenta con poca bibliografía para estas pruebas, es posible definir parámetros de desempeño que el laboratorio debe tener en cuenta para poder liberar resultados confiables y de utilidad clínica. Tanto el control de calidad interno como el externo son dos valiosas herramientas orientadas a cumplir con la misión de los laboratorios en la cual el objetivo principal es la seguridad del paciente. Desde el Programa de Evaluación Externa de la Calidad se ofrecen varios subprogramas para el control de pruebas serológicas, que proporcionan a cada participante una evaluación objetiva del comportamiento de su laboratorio, información de los métodos disponibles y la identificación de factores que puedan afectar sus resultados.

Although the quality control in quantitative determinations has wide dissemination, an extensive bibliography that supports it and it is constantly being updated, this is not the case when referring to qualitative tests such as those used in serological diagnoses. Before using this serological test in the laboratory, it is important to have information about its characterization, usefulness and more convenient working algorithms. Although there is little literature for these tests, it is possible to define performance parameters that the laboratory must take into account to be able to release reliable and clinically useful results. Both internal and external quality control are valuable tools aimed at fulfilling the mission of laboratories in which the main objective is patient safety. From the External Quality Assessment Programme, several subprogrammes are offered for the control of serological tests, providing each participant with an objective evaluation of the behaviour of their laboratory, information on the available methods and the identification of factors that can affect the results.

Embora o controle de qualidade em determinações quantitativas possua ampla divulgação e haja extensa bibliografia que o apoie e esteja em constante atualização, o mesmo não acontece quando se refere a testes qualitativos, como os utilizados em diagnósticos sorológicos. Antes de usar esse teste sorológico em laboratório, é importante ter informações sobre sua caracterização, utilidade e algoritmos de trabalho mais convenientes. Embora exista pouca literatura para esses testes, é possível definir parâmetros de desempenho que o laboratório deve levar em consideração para poder liberar resultados confiáveis e clinicamente úteis. O controle de qualidade interno e externo são duas ferramentas valiosas destinadas a cumprir a missão dos laboratórios na qual o objetivo principal é a segurança do paciente. A partir do Programa de Avaliação Externa da Qualidade, são oferecidos vários subprogramas para o controle de testes sorológicos, proporcionando a cada participante uma avaliação objetiva do comportamento de seu laboratório, informações sobre os métodos disponíveis e identificação de fatores que possam afetar os resultados.

Efectividad de los métodos diagnósticos para la detección de ehrlichiosis monocítica humana y canina / Effectivity of diagnostic methods for the detection of human and canine monocytic ehrlichiosis

Resumen La ehrlichiosis es una enfermedad transmitida por la picadura de garrapatas que afecta a perros y humanos, causada por las especies Ehrlichia canis y E. chaffeensis, respectivamente. Estas bacterias son gramnegativas, intracelulares obligadas, de aspecto cocoide a pleomorfo, que infectan los monocitos y desencadenan síntomas como fiebre elevada, anorexia, trombocitopenia, hemorragias, anemia y problemas graves como esplenomegalia, hepatomegalia y meningitis. Para diagnosticar esta enfermedad existen diversos métodos, entre los que se encuentran los hematológicos que evalúan la morfología de los monocitos en búsqueda de mórulas y la serología, que incluye la búsqueda de anticuerpos anti-Ehrlichia, pero que se encuentra limitado debido a la reactividad cruzada que presenta. Por otra parte, el cultivo de especies de Ehrlichia ha resultado ser un método efectivo para la obtención de antígenos y así desarrollar ensayos por inmunofluorescencia indirecta (IFI). El método por reacción de polimerasa en cadena ofrece un diagnóstico definitivo por tener una mayor sensibilidad y especificidad que los otros métodos, al haberse desarrollado cebadores género-específicos, así como especie-específicos. En esta revisión, se discutirán los diversos métodos aplicados al diagnóstico de esta enfermedad, así como las ventajas y desventajas que estos presentan.

Ehrlichiosis is a disease transmitted by tick's bite that affect dogs and humans caused by the species Ehrlichia canis and E. chaffeensis, respectively. These bacteria are obligated intracellular gram negatives, with a cocoid to pleomorph aspect and can infect monocytes and trigger symptoms such as high fever, anorexia, thrombocytopenia, hemorrhages, anemia, and some serious problems such as splenomegaly, hepatomegaly and meningitis. There are several diagnostic tests for ehrlichiosis such as the hematological ones that evaluate the morphology of the monocytes in search of morulae; serological tests that includes the search of anti-Ehrlichia antibodies, although they might be limited due to cross reaction with other species. In other hand, the culture of Ehrlichia species is an effective method to obtain antigens and even develop indirect immunofluorescence assays (IFA). The polymerase chain reaction offers a definitive diagnosis associated to the use of genus-specific and species-specific primers, as well as its increased sensibility and specificity, compared to the others methods. Thus, in this review, we will discuss various methods applied to the diagnosis of this disease, as well as the advantages and disadvantages that these present.

Manifestações oculares na febre Chikungunya / Ocular Manifestations of Chikungunya Fever

Resumo A febre Chikungunya é um problema de saúde pública mundial, com potencial para gerar epidemias de alta morbidade, visto que elevado número de pacientes pode apresentar sequelas articulares prolongadas e alterações oftalmológicas. As manifestações oftalmológicas podem estar presentes na fase aguda da doença ou ter início após várias semanas da instalação do quadro. Na literatura mundial é descrito desde alterações mais comuns e de fácil tratamento como conjuntivites até alterações mais complexas e que podem cursar com sequelas visuais graves como a retinite e neurite óptica.

Abstract Chikungunya fever is a world public health problem with the potential to generate epidemics of high morbidity, since a high number of patients may present prolonged joint sequelae and ophthalmological alterations. Ophthalmologic manifestations may be present in the acute phase of the disease or begin after several weeks of the onset of the disease. In the world literature is described from more common and easy to treat changes such as conjunctivitis to more complex changes and that can occur with severe visual sequelae such as retinitis and optic neuritis.

Leprosy: prevalence and factors associated with seropositivity for anti-NDO-LID antibodies in children under 15 years of age

Abstract: Background: A high prevalence of leprosy among children under 15 years of age indicates the need to implement actions to prevent new cases of the disease. Serological tests have been developed with the aim of helping to control the disease by indicating, through seropositivity, the presence of infection. Objective: To analyze the prevalence and factors associated with seropositivity rate for anti-NDO-LID antibodies in children under 15 years of age, contacts of leprosy patients. Method: We performed a cross-sectional study with 210 children under 15 years old of age. Of them, 50 were household contacts and 160 were neighborhood contacts living in the municipality of Cuiabá, state of Mato Grosso, in 2016. The data were obtained from interviews and the NDO-LID rapid test during home visits from February to July 2016. For the analysis, we used Poisson regression and prevalence ratio. Results: Seropositivity in contacts was 6.2%. Variables associated with seropositive tests included sex (PR = 1.05; 95% CI: 1.01 - 1.08), race/skin color (PR = 0.95; 95% CI: 0.90 - 0.99), residence area (PR = 1.05; 95% CI: 1.01 - 1.09), and number of people per household (PR = 1.06; 95% CI: 1.02 - 1.08). Study Limitations: The small sample size, besides leading to wide confidence intervals, may have been a limitation for the identification of associated factors. Conclusions: The prevalence of seropositivity was high. Variables associated with NDO-LID seropositivity included female sex, not to be brown skinned, live in urban areas, and live with five or more people.

Características clínicas y epidemiológicas de pacientes con paracoccidioidomicosis diagnosticados en un hospital de Resistencia, Chaco / Clinical and epidemiological characteristics of patients with paracoccidioidomycosis diagnosed in a hospital of Resistencia, Chaco

Con el objetivo de describir las características clínico-epidemiológicas de la paracoccidioidomicosis, se realizó un estudio descriptivo de los casos diagnosticados por el Servicio de Microbiología Clínica del hospital de adultos Dr. Julio C. Perrando, de la ciudad de Resistencia (Chaco, Argentina). Entre 2011 y 2014 se detectaron 46 casos. En 2013 y 2014 se constató un incremento de la tasa de incidencia de alrededor de 4 veces con respecto a los anos anteriores. La forma crónica fue la predominante, con una media de edad de los pacientes de 53 anos. Del total de ellos, a 39 se les realizaron pruebas serológicas. En 15 de 39 casos, las pruebas serológicas fueron la única herramienta diagnóstica, mientras que en 4 de estos casos con diagnóstico microbiológico, la prueba resultó no reactiva. La inclusión de la paracoccidioidomicosis en el diagnóstico diferencial de pacientes de áreas endémicas que presentan un síndrome infeccioso inespecífico y la aplicación de las herramientas diagnósticas disponibles contribuyen al diagnóstico oportuno, así como a disminuir las secuelas de esta afección y su impacto socioeconómico.

In order to describe the clinical and epidemiological characteristics of paracoccidioidomycosis, a descriptive study of all the cases diagnosed by the Clinical Microbiology Service at Dr. Julio C. Perrando hospital in the city of Resistencia (Chaco Province, Argentina) was conducted. Between 2011 and 2014, 46 cases were detected. In the period 2013-2014, an almost 4-fold increase in the incidence rate was detected. The chronic form of the disease was predominant with an average age of 53 years. Serological tests in 39 out of 46 patients were performed. In 15 of 39 patients, serological tests were the only diagnostic tool while in 4 patients with a microbiological diagnosis serological tests were non-reactive. In patients from endemic areas with non-specific infectious syndrome it is important to include paracoccidioidomycosis in the differential diagnosis and to apply all available diagnostic tools to reach a timely diagnosis and to reduce the long-term sequelae and their socio-economic impact.

Evaluación del sistema Brucellacapt® para el diagnóstico serológico de la brucelosis humana en Cuba / Evaluation of the system Brucellacapt® for serological diagnosis of human brucellosis in Cuba

Introducción: En el diagnóstico microbiológico de la brucelosis, los métodos serológicos son los más utilizados. Brucellacapt® ofrece la ventaja de detectar en cualquier momento de la enfermedad anticuerpos aglutinantes y no aglutinantes contra Brucellaspp. Objetivos: Evaluar y aplicar el sistema serológico comercial Brucellacapt® para la detección de anticuerpos contra Brucella spp. Métodos: Se realizó una investigación en servicios y sistemas de tipo observacional con un estudio de caso control anidado, en el periodo de enero de 2015 a junio de 2016, en el Laboratorio Nacional de Referencia de Espiroquetas y Brucelas. Se evaluaron 50 sueros de casos y 100 de controles por Brucellacapt®. Se aplicó Brucellacapt® en 695 sueros de casos con sospecha clínica o epidemiológica de la enfermedad, los cuales fueron confirmados por ELISA. Resultados: Brucellacapt® mostró 100 por ciento de sensibilidad, 83 por ciento de especificidad, 54,6 por ciento de reactividad y 16,4 por ciento de positividad en las muestras estudiadas. Se confirmó por ELISA la presencia de anticuerpos IgM (21, 2 por ciento), IgG (6,1 por ciento) e IgM + IgG (7,6 por ciento). Doscientas cuarenta y seis muestras y otras siete fueron reactivas y positivas respectivamente, solo por Brucellacapt®. Conclusiones: Brucellacapt® contribuyó a la detección de anticuerpos en los sueros de pacientes sospechosos de la enfermedad con valores aceptables de sensibilidad y especificidad diagnóstica. Este resultado sugiere su implementación en la red nacional de laboratorios cubanos para fortalecer el diagnóstico y la vigilancia de la brucelosis humana en Cuba(AU)

Introduction: Serological methods are the most commonly used for the microbiological diagnosis of brucellosis. Brucellacapt® has the advantage of detecting agglutinating and non-agglutinating antibodies against Brucella spp. at any point in the evolution of the disease. Objectives: Evaluate and apply the commercially available serological system Brucellacapt® for detection of antibodies against Brucella spp. Methods: A nested case-control observational study was conducted of services and systems from January 2015 to June 2016 at the Spirochaete and Brucella National Reference Laboratory. Evaluation was performed of 50 serum samples from cases and 100 from controls using Brucellacapt®. The system was also used in 695 serum samples from clinically or epidemiologically suspected cases, which were confirmed by ELISA. Results: Brucellacapt® showed 100 percent sensitivity, 83 percent specificity, 54.6 percent reactivity and 16.4 percent positivity in the samples studied. Presence of the following antibodies was confirmed by ELISA: IgM (21.2 percent), IgG (6.1 percent) and IgM + IgG (7.6 percent). Two hundred forty-six samples and another seven were reactive and positive, respectively, only by Brucellacapt®. Conclusions: Brucellacapt® contributed to antibody detection in serum samples from suspected cases, with acceptable diagnostic sensitivity and specificity values. This result suggests its implementation in the Cuban national network of laboratories to strengthen the diagnosis and surveillance of human brucellosis in Cuba(AU)

Development and standardization of a western blotting test for detection of antibodies against B. abortus / Desenvolvimento e padronização do teste Western blotting para detecção de anticorpos contra B. abortus

Brucellosis is an infectious disease caused by bacteria of the genus Brucella spp. with diagnosis based on use of serological techniques. The present study aimed to develop and standardize a western blotting (WB) test for detection of antibodies against B. abortus. Samples from two groups of cattle were analyzed: group I: 60 serum samples from true positive and true negative vaccinated animals (30 positive samples from infected animals according to rose bengal test (RBT), 2-mercaptoethanol, serum agglutination test (SAT) and complement fixation test (CFT) and 30 RBT negatives samples); group II: 383 field samples (90 positive and 293 CFT negative sera). The most reactive band in the western blotting, which properly identified and separated infected from non - infected had a molecular weight of ≤ 20kDa. The sensitivity, specificity and accuracy of the WB compared to RBT was 93%, 99%, 98%, respectively and k= 0.938. When compared to CFT, the sensitivity, specificity and accuracy of the WB was 97%, 98% and 97%, respectively and k= 0.929. The WB developed and standardized in the present study is a serological test with potential use as a confirmatory test for the diagnosis of bovine brucellosis.(AU)

A brucelose é uma doença infectocontagiosa, causada por bactérias do gênero Brucella spp., com diagnóstico baseado no emprego de técnicas sorológicas. Objetivou-se neste estudo desenvolver e padronizar um teste Western blotting (WB) para detecção de anticorpos contra B. abortus. Foram analisados dois grupos de amostras bovinas: grupo I, com 60 amostras de animais verdadeiros positivos e verdadeiros negativos vacinados (30 amostras positivas de animais infectados e positivos nos testes de antígeno acidificado tamponado (AAT), 2 - mercaptoetanol (2 - ME), soroaglutinação lenta em tubos (SAT) e fixação do complemento e de 30 amostras negativas no AAT); grupo II, com 383 amostras de campo, sendo 90 soropositivas e 293 soronegativas no TFC. O resultado da análise do WB revelou peso molecular ≤20kDa como sendo a área mais reativa e característica para identificação e separação dos animais infectados dos não infectados. A sensibilidade, a especificidade e a acurácia do WB, quando este foi comparado com o AAT, foram, respectivamente, 93%, 99% e 98%, e k= 0,938. Quando comparadas com a TFC, a sensibilidade, a especificidade e a acurácia foram 97%, 98% e 97%, respectivamente, e k= 0,929. O WB padronizado neste estudo mostrou-se um teste sorológico com potencial uso como teste confirmatório no diagnóstico da brucelose bovina.(AU)

Fatores associados ao não uso de preservativo e prevalência de HIV, hepatites virais B e C e sífilis: estudo transversal em comunidades rurais de Ouro Preto, Minas Gerais, entre 2014 e 2016 / Factores asociados al hábito de no uso del preservativo y prevalencia del VIH, hepatitis virales B y C y sífilis: estudio transversal en comunidades rurales de Ouro Preto, Minas Gerais, Brasil, 2014-2016 / Factors associated with non-use of condoms and prevalence of HIV, viral hepatitis B and C and syphilis: a cross-sectional study in rural communities in Ouro Preto, Minas Gerais, Brazil, 2014-2016

Objetivo: analisar os fatores sociodemográficos relacionados ao não uso do preservativo nas relações sexuais e a prevalência de infecções sexualmente transmissíveis (IST) em comunidades rurais de Ouro Preto, Minas Gerais, Brasil, entre 2014 e 2016. Métodos: foram coletados dados de entrevista individual e realizados testes rápidos; as associações foram testadas pelo modelo de regressão de Poisson, com intervalo de confiança de 95% (IC95%). Resultados: foram detectados 3,8 casos/10 mil habitantes de hepatite B e sífilis, e 1,3/10 mil hab. de hepatite C; não foram detectados casos de HIV; na análise multivariada, foram encontradas maiores prevalências de não uso de preservativos entre indivíduos casados/em união estável/viúvos (RP=1,20 - IC95% 1,06;1,36). Conclusão: o grupo com maior prevalência de não uso de preservativo é o de pessoas com relacionamento fixo; novos casos de sífilis e de hepatites virais foram detectados pelo teste rápido, aplicado no inquérito.

Objetivo: investigar el perfil sociodemográfico asociado al no uso del preservativo en las relaciones sexuales y describir la prevalencia de infecciones de transmisión sexual (ITS) en comunidades rurales de Ouro Preto, Minas Gerais, Brasil, entre 2014 y 2016. Métodos: se recolectaron datos en entrevista individual y se realizaron pruebas rápidas; las asociaciones fueron analizadas por la regresión de Poisson, con intervalo de confianza de 95% (IC95%). Resultados: se detectaron 3,8 casos/10 mil habitantes de hepatitis B y sífilis, y 1,3/10 mil habitantes de hepatitis C; no se detectaron casos de VIH; el análisis multivariado mostró mayor prevalencia de no uso del preservativo para personas casadas /en unión estable/viudos (RP=1,20 - IC95% 1,06;1,36). Conclusión: se detectaron nuevos casos de sífilis y hepatitis virales por las pruebas rápidas en esta investigación; el grupo de mayor riesgo de no usar preservativo fue de personas casadas/con relación fija.

Objective: to investigate socio-demographic factors associated with non-use of condoms, and to describe the prevalence of sexually transmitted infections (STI) in rural communities of Ouro Preto, Minas Gerais, Brazil, 2014 to 2016. Methods: data were gathered from individual interviews and rapid tests were performed; associations were tested using Poisson regression, with a 95% confidence interval (95%CI). Results: we detected 3.8 cases/10,000 inhabitants for hepatitis B and syphilis, and 1.3 cases/10,000 inhabitants for hepatitis C; no HIV cases were detected; in the multivariate analysis we found higher prevalence rates of condom non-use among the group of individuals who were married, had common law partners or were widowed (PR=1.20 - 95%CI 1.06;1.36). Conclusion: individuals in a stable relationship formed the group with the highest prevalence rate of condom non-use; new syphilis and viral hepatitis cases were detected using rapid tests during the survey.