Differences in the replicative capacities of clinical isolates of dengue virus in C6/36 cells and in urban populations of Aedes aegypti from Colombia, South America

ABSTRACT Dengue, the most prevalent arboviral disease worldwide, is caused by any of the four dengue virus (DENV) serotypes that co-circulate constantly in hyperendemic areas such as Medellin (Colombia), and these serotypes are transmitted by mosquitoes of the genus Aedes. In this study, we evaluated the replicative capacity of strains isolated in Medellin between 2003 and 2007 in C6/36 cells and in colonies of Aedes aegypti collected during 2010-2011 from high or low-incidence areas within the same city. The phylogenetic analysis grouped isolates according to the predominant genotypes found in the Americas, and the in vitro characterization showed differences in the morphological changes induced by the isolates of each of the isolated serotypes compared to the reference serotypes. In vitro replicative capacity studies demonstrated that genomic copy number increased at four days post-infection and that cell viability decreased significantly compared to the control for all serotypes. The largest number of genomic copies in C6/36 was produced by DENV-2, followed by DENV-1 and DENV-4; DENV-3 produced the smallest number of genomic copies and had the smallest negative effect on cell viability. Finally, differences in the in vivo replication of intercolonial serotypes between the Rockefeller colony and the field colonies and among the intracolonial serotypes were found. The replication of DENV-2 at 7 and 14 days in both high- and low-incidence colonies was higher than that of the other serotypes, and replication of DENV-3 in the mosquito colonies was the most stable on the days evaluated. Our results support the notion that replication and, possibly, DENV transmission and severity depend on many factors, including serotype and vector characteristics.

Actualización en diagnóstico del dengue: evolución de las técnicas y su aplicación real en la clínica / Update on dengue diagnosis: techniques evolution and their clinical application

Resumen: El dengue es la enfermedad viral transmitida por vectores de mayor importancia en el mundo. Debido a que hasta el momento no existe una vacuna licenciada para la prevención de la infección ni una terapia específica para controlar la enfermedad, el diagnóstico temprano y específico resulta ser una herramienta de vital importancia para brindar un tratamiento rápido y oportuno al paciente. Aunque hace más de seis décadas se dispone de una variedad de técnicas de laboratorio para el diagnóstico de esta enfermedad, es frecuente encontrar que noexista un consenso en el personal o en las entidades colombianas encargadas del diagnóstico de laboratorio sobre las ventajas y las limitaciones de estas técnicas, lo que dificulta dichodiagnóstico. Adicionalmente, debido a que la mayoría de técnicas que se usan se basan en la respuesta inmune frente al virus, y se pueden presentar reacciones cruzadas con otros virusgenéticamente relacionados, la posibilidad de falsos positivos es alta. Principalmente, por estarazón, en los últimos años ha aumentado el desarrollo y la validación de técnicas moleculares,ya que son más sensibles, específicas y rápidas que las técnicas celulares e inmunológicas.Teniendo en cuenta estos antecedentes, se hace necesario comparar a la luz de la literatura disponible la utilidad real de las técnicas de laboratorio existentes, clasificándolas en celulares,inmunológicas y moleculares.

Abstract: Dengue is the vector-borne viral disease of major importance in the world. Because so far there is no licensed vaccine that can be used to prevent the infection and there is no specific antiviral treatment, the early and specific diagnosis is a powerful tool to provide prompt and timely management of the patient. Despite during more than six decades there has been available a wide variety of laboratory techniques that can be used for the diagnosis of dengue, there is often lack of a consensus among the people or entities responsible of the diagnosis about the advantages and disadvantages of these techniques; for this reason, the diagnosis of dengue is more difficult. Moreover, taking into account that most of the techniques used are based on the immune response against dengue virus, and a cross reactivity with other viruses genetically related could be possible; the possibility of false positives is high. Accordingly, in recent years the development and validation of molecular techniques has increased, as they are more sensitive, specific and rapid than the cellular and immunological techniques. Therefore, it is necessary to compare the usefulness of laboratory techniques to dengue diagnosis, such as cellular, immunological and molecular tests.

Competencia vectorial: consideraciones entomológicas y suinfluencia sobre la epidemiología del dengue / Vector competence: entomological considerations and its implications on the epidemiology of dengue

Las enfermedades transmitidas por vectores constituyen un complejo problema de salud en el ámbito mundial, especialmente en áreas tropicales y subtropicales, que por sus condiciones ambientales favorecen la supervivencia del vector. El dengue es la principal enfermedad transmitida por vectores en el mundo, cuyo agente etiológico es el Virus Dengue, el cual es transmitido por la picadura de mosquitos hematófagos de la especie Aedes aegypti. La interacción virus-vector es esencial para una transmisión efectiva y depende de factores tanto virales como vectoriales. Entre los factores relacionados con el vector, la competencia vectorial es considerada de gran importancia, pues se refiere a la capacidad intrínseca del vector para infectarse con el virus, permitir su replicación y posteriormente su transmisión a un huésped susceptible. A su vez, la habilidad para ser un buen vector depende principalmente de barreras naturales a la infección, barreras inmunológicas y presencia de receptores específicos para el virus. Todas estas características, sumadas a las características propias del virus, favorecerán o no la transmisión del mismo. Un estudio integral dela relación virus-vector permitirá comprender sus implicaciones en la epidemiología de la enfermedad. Estos temas han sido revisados en el presente documento, discutiendo su importancia dentro del contexto entomológico y epidemiológico.

Vector-borne diseases are a serious problem in public health, especially in tropical and subtropical areas where environmental conditions favor the survival and expansion of vectors into new habitats. Dengue fever is one of the most important vector-borne diseases transmitted by arthropods (arbovirosis) worldwide. The etiologic agent of the disease is the dengue virus, which is transmitted by the bite of bloodsucking Aedes aegypti mosquitoes. The virus-vector interaction is essential for the efficient transmission of the disease, and depends on both, viral factors and vector competence or intrinsic vector capacity to be infected with the virus. Vector competence allows virus replication and subsequent transmission to susceptible hosts. Factors that influence vector capacity include: natural barriers to infection, immunological defenses as well as the presence of receptors for the virus. All these factors inaddition to the viral characteristics will determine the degree of transmission. There is a need for a better understanding of the virus-vector relationship and its epidemiological implications. These issues are addressed in this article.