Padronização da detecção de Zika virus em Aedes aegypti e Aedes albopictus usando cartões FTA em condições de laboratório

• Ano de defesa: 2019
• Autor(a) principal: Bruna Antoniety Oliveira
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Minas Gerais; Brasil; ICB - DEPARTAMENTO DE PARASITOLOGIA; Programa de Pós-Graduação em Parasitologia; UFMG
• Palavras-chave em Português: Parasitologia; Zika virus; Aedes; RNA Viral; Preservação de Amostras; FTA Card; DNA Viral
• Link de acesso: http://hdl.handle.net/1843/54305

Zika virus (ZIKV) is an arbovirus that has been causing worldwide concern since the major outbreaks that occurred in Brazil and worldwide. In 2015, with the entry of the virus in Brazil, numerous cases of ZIKV-associated congenital syndrome (microcephaly) were registered. At present, there is no vaccine available for ZIKV and for this reason the only way to prevent the disease is vector control. Currently, traps to capture adult vectors are used as a method for monitoring insect populations and the subsequent analysis of the genetic material of these vectors is used as viral monitoring. The Flinders Technology Associates (FTA) cards are applied to preserve genetic material from biological samples and have being studied as an alternative for viral monitoring in Aedes spp. Therefore, the present study aimed to evaluate the use of FTA card in the collect of saliva and excreta from Ae. aegypti and Ae. albopictus infected for ZIKV detection under laboratory conditions. For evaluation of ZIKV detection from saliva and excreta, mosquitoes were artificially infected with blood containing ZIKV culture and separated into pots containing groups of 1, 3 and 5 specimens. Each pot contained a card for collection of excreta and mosquitoes were placed in contact with FTA cards impregnated with a 10% solution of manuka honey for 48 h for saliva release, 14 days after blood feeding. Subsequently, cards containing saliva and excreta were subjected to molecular analysis to identify viral genetic material. The results showed that it was possible to store and identify viral genetic material after 15 days of preservation in the card of samples from saliva and excreta extracted using a commercial kit. It was also possible to detect viral RNA from saliva of ZIKV-infected mosquitoes from the card, showing the ability to store and preserve viral RNA even in small amounts. The excreta analysis in cards has been shown to be a sensitive method for detection of viral RNA. It is believed that this technique will facilitate the process of collection and preservation of genetic material, detection and viral monitoring under field conditions, allowing an estimation of the risk of transmission.