Development and characterization of serotype-specific monoclonal antibodies against Dengue virus NS1 / 生物工程学报

Dengue virus (DENV) is the most widely transmitted arbovirus in the world. Due to the lack of diagnostic technology to quickly identify the virus serotypes in patients, severe dengue hemorrhagic fever cases caused by repeated infections remain high. To realize the rapid differential diagnosis of different serotypes of DENV infection by immunological methods, in this study, four DENV serotype NS1 proteins were expressed and purified in mammalian cells. Monoclonal antibodies (MAbs) against NS1 protein were obtained by hybridoma technology after immunizing BALB/c mice. Enzyme-linked immunosorbent assay, indirect immunofluorescence assay, dot blotting, and Western blotting were used to confirm the reactivity of MAbs to viral native NS1 and recombinant NS1 protein. These MAbs include not only the universal antibodies that recognize all DENV 1-4 serotype NS1, but also serotype-specific antibodies against DENV-1, DENV-2 and DENV-4. Double antibody sandwich ELISA was established based on these antibodies, which can be used to achieve rapid differential diagnosis of serotypes of DENV infection. Preparation of DENV serotype-specific MAbs and establishment of an ELISA technology for identifying DENV serotypes has laid the foundation for the rapid diagnosis of DENV clinical infection.

Anticuerpos policlonales contra la proteína recombinante NS3 del virus del dengue / Polyclonal antibodies against recombinant dengue virus NS3 protein

Resumen Introducción: El dengue es una enfermedad causada por uno de los cuatro serotipos del virus del dengue (DENV) y es endémica en, aproximadamente, 130 países. Su incidencia ha aumentado notablemente en las últimas décadas, así como la frecuencia y la magnitud de los brotes. A pesar de los esfuerzos, no existen tratamientos profilácticos ni terapéuticos contra la enfermedad y, en ese contexto, el estudio de los procesos que gobiernan el ciclo de infección del DENV es esencial para desarrollar vacunas o terapias antivirales. Una de las moléculas del DENV más prometedoras es la proteína no estructural 3 (NS3), la cual es indispensable para la replicación viral y es uno de los principales blancos inmunológicos durante la infección. Objetivo: Producir anticuerpos policlonales para contribuir a los futuros estudios sobre las interacciones entre la proteína NS3 y otras proteínas celulares. Materiales y métodos: Se expresaron dos proteínas recombinantes del dominio helicasa de NS3 del DENV de serotipo 2, las cuales se emplearon para inmunizar ratas y producir anticuerpos policlonales. Resultados: Los anticuerpos producidos fueron útiles en ensayos de Western blot e inmunofluorescencia y se reportó por primera vez un anticuerpo policlonal anti-NS3 que permitió la inmunoprecipitación de la proteína viral y la detecta con Western blot sin necesidad de inducir sobreexpresión de NS3 o de usar extractos de células marcados metabólicamente con radioisótopos. Conclusión: Las proteínas recombinantes expresadas y los anticuerpos producidos constituyen herramientas valiosas para estudiar procesos infecciosos del DENV que involucren a la proteína NS3 y evaluar pruebas dirigidas a interferir las funciones de esta proteína.

Abstract Introduction: Dengue is a disease caused by one of four serotypes of the dengue virus (DENV) and is endemic in approximately 130 countries. The incidence of dengue has increased dramatically in recent decades, as well as the frequency and magnitude of outbreaks. Despite all efforts, there are no prophylactic or therapeutic treatments for the disease. Accordingly, research on the processes governing the DENV infection cycle is essential to develop vaccines or antiviral therapies. One of the most attractive DENV molecules to investigate is nonstructural protein 3 (NS3), which is essential for viral replication and a major immune target for infection. Objective: To produce antibodies to support future studies on NS3 and its cellular interactions with other proteins. Materials and methods: Two recombinant proteins of the helicase domain of DENV NS3 serotype 2 were expressed, and used to immunize mice and produce polyclonal antibodies. Results: The antibodies produced were useful in Western blot and immunofluorescence tests. We report an NS3 antibody that immunoprecipitates the viral protein and detects it in Western blot with no need to over-express it or use cell extracts with metabolic radiolabeling.

La Escuela de Salud Pública de México: innovación educativa y tecnológica en el nuevo milenio / School of Public Health of Mexico: Educational and technological innovation in the new millennium

Este artículo fue concebido para analizar la función de la Escuela de Salud Pública de México (ESPM) desde el año 2000 hasta el presente. Uno de sus puntos centrales es el análisis del proceso de reorientación de la labor educativa de la escuela con la finalidad de responder a los retos en materia de salud y educación surgidos a finales del siglo XX. Para exponer cómo ha evolucionado dicho proceso, retomamos tres ejes rectores que caracterizan la labor de la escuela en la actualidad: el cambio de modelo pedagógico, la incorporación de las tecnologías de la información y las comunicaciones, y la profesionalización de la docencia. Con la exposición de este tema, y a través del contraste entre el pasado y el presente, buscamos completar la historia de trabajo ininterrumpido de la Escuela durante sus 92 años de existencia, que ha trascendido los confines del país.

This article was conceived to analyze the work of the School of Public Health of Mexico (ESPM for is acronym in Spanish) from the year 2000 to the present day. One of the highlights that we will examine is the reorientation of the educational work of the school in order to meet the challenges in health and education that emerged during the end of the twentieth century. In order to explain the evolution of this process, we will describe the three main guiding principles that characterize the present work of the school: the pedagogical model's change, the incorporation of the information and communication technologies, and the professionalization in teaching. The purpose of this work is to define those guiding principles, and to expose, through the contrast between past and present, the complete history of uninterrupted work of the School of Public Health of Mexico during its ninety-two years of existence, that has gone beyond the boundaries of the country.

Epidemiological Investigation of an Outbreak of Hepatitis A at a Residential Facility for the Disabled, 2011 / 예방의학회지

OBJECTIVES: An outbreak of hepatitis A occurred at a residential facility for the disabled in July 10, 2011. This investigation was carried out to develop a response plan, and to find the infection source of the disease. METHODS: A field epidemiologist investigated the symptoms, vaccination histories, living environments, and probable infection sources with 51 residents and 31 teachers and staff members. In July 25, 81 subjects were tested for the hepatitis A virus antibody, and specimens of the initial 3 cases and the last case were genetically tested. RESULTS: Three cases occurred July 10 to 14, twelve cases August 3 to 9, and the last case on August 29. Among the teachers and staff, no one was IgM positive (on July 25). The base sequences of the initial 3 and of the last case were identical. The vehicle of the outbreak was believed to be a single person. The initial 3 patients were exposed at the same time and they might have disseminated the infection among the patients who developed symptoms in early August, and the last patient might have, in turn, been infected by the early August cases. CONCLUSIONS: The initial source of infection is not clear, but volunteers could freely come into contact with residents, and an infected volunteer might have been the common infection source of the initial patients. Volunteers' washing their hands only after their activity might be the cause of this outbreak. Although there may be other possible causes, it would be reasonable to ask volunteers to wash their hands both before and after their activities.

Does Rota virus infection cause persistent diarrhoea in childhood?

Rota virus is the commonest agent for infectious diarrhoea in childhood. Whether an acute diarrhoea episode due to Rota virus constitutes a risk factor for persistence of diarrhoeal episode has not been well evaluated. This study aimed at evaluating the role of antecedent Rota virus infection in the causation of persistent diarrhoea. One hundred twenty children with acute watery diarrhoea were investigated for evidence of recent Rota virus infection and were followed up for 2 weeks to determine the proportion developing persistent diarrhoea. Rota virus antigen in stool and IgM class antibodies in serum were detected in 33 (27.5%) of the cases, only 2 (6.06%) of whom developed persistent diarrhoea. On the other hand 11.3% children without evidence for antecedent Rota virus infection developed persistent diarrhoea. The prevalence of Rota virus antigen in stools of children who developed persistent diarrhoea was 16.7%. In another group of 25 children with persistent diarrhoea definite evidence for a Rota virus infection was detected in only 2 (8%) cases. It was therefore concluded that antecedent Rota virus infection does not increase the risk for abnormal prolongation of an episode of acute diarrhoea.