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Cuantificación relativa de genes relacionados con la respuesta inmune antiviral en una población venezolana de Aedes (stegomyia) aegypti

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Author(s): Sergio Montaner | Zoraida Fernández Zoraida | Angélica Jiménez | Johanny Ruiz | Marifel Carrozza | José Rivero | Flor Herrera

Journal: Salus Online
ISSN 1316-7138

Volume: 15;
Issue: 3;
Start page: 39;
Date: 2011;
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Keywords: Dengue | PCR | RNA interference | Aedes aegypti

ABSTRACT
DENV-3 is the responsible for the second major percentage of hemorrhagic severe affections caused by this virus, only overcome by DENV-2. The virulence of this serotype is the cause of broad scale fever outbreaks in different countries, especially in Venezuela where it circulates actively throughout the year. This is the main reason of investigative developments in the virus-vector interactions field to find aneffective epidemic control. In this study, we evaluated the expression of dcr-2 and ago-2, two RNAi antiviral pathway genes in mosquitoes to study the fold change in the expression of these genes in the mosquito vector Aedes (stegomyia) aegypti infected with DENV-3. We artificially infected a population of mosquitoes from Trujillo state (Venezuela) and prepared three (3) experimental groups (15dpi, 20dpi anda control group). Later, we isolated, quantified and applied a reverse transcription to the total RNA obtained of mosquitoes and the infection in the experimental groups was detected performing a 2% agarose gel electrophoresis of the PCR products of the total RNA (511bp for DENV and 290bp for DENV-3). In addition, the infection in the experimental groups was confirmed by the detection of PCR products.The fold change in dcr-2 and ago-2 expression genes was quantify by Real Time PCR. As results, the fold change expression in the ago-2 and dcr-2 were equal to the non-infected mosquitoes “control group” (p

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