Investigation of alphaviruses in hematophagous insects

Abstract

Arboviruses are viral infections transmitted by arthropods (mosquitoes), caused by viruses of several genera, among them the Alphavirus, belonging to the family Togaviridae. Among the species of alphavirus that affect the countries of the American continent and which are of medical importance, we have: the Mayaro Fever virus, the Chikungunya Fever virus and the Equine Encephalitis virus. Molecular biology is widely used as a method of viral isolation in vectorsand among the diverse techniques available for the identification of Arboviruses, the reverse transcriptase - polymerase chain reaction (RT/PCR) is one of the most widely used. The most commonly used method of viral vector RNA extraction involves maceration of the insect, damaging the morphology and making it impossible to identify the vector. In order to integrante molecular and morphological studies, the enzymatic method of genetic material extraction in arthropods, developed by our research group, allows the identification of genética material of pathological agents and vertebrate present in The insect, in addition tô enabling morphological and molecular identification of the insect studied. This project aims to detect alphavirus present in the New World in Diptera insects collected in public parks in the ABC region of São Paulo (Santo André and São Bernardo do Campo) and in the city of Marília. The applied methodology will involve the collection and identification of the insects present in the regions included in the study, the RNA extraction of the insects through the enzymatic method with proteinase K followed by The diagnosis of alphavirus by RT/PCR, the identification of the origin of the blood presente in the hematophagous insects by PCR and the taxonomic identification of alphavirus positive insects. PCR detection will be performed using primers described in the literature for Chikungunya, Mayaro and vertebrate animals, including humans. (AU)