Sequencing of Flavivirus in mosquitoes (Diptera:Culicidae) collected in the city of São Paulo.

• Grant number: 23/12892-5
• Support Opportunities: Scholarships in Brazil - Master
• Effective date (Start): November 01, 2024
• Effective date (End): November 30, 2025
• Field of knowledge: Health Sciences - Collective Health - Public Health
• Principal Investigator: Licia Natal Fernandes
• Grantee: Gabriel Matheus do Nascimento
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Several epidemics caused by flaviviruses have already been recorded in Brazil, including dengue fever, yellow fever and Zika fever.In the country, in addition to flaviviruses of medical importance, there is evidence of circulation of insect-specific flaviviruses (ISFs), a group known to replicate only in insect cells, such as Cell fusing agent virus (CFAV), Culex flavivirus (CxFV), Aedes flavivirus (AEFV) and others.There is a hypothesis that a mosquito infected by an insect-specific flavivirus may be resistant to a second infection caused by medically important flaviviruses due to the superinfection exclusion mechanism, which arouses interest in the group, due to the potential impact on the vector competence of mosquitoes.Although there are several reports about the occurrence of these viruses in mosquitoes, little is known about them.The use of Next Generation Sequencing techniques has generated a lot of data on ISFs, both by discovering new viruses and by increasing the number and size of sequences generated.Considering the importance of Flaviviruses, the objective of the present work will be to sequence the complete genome of Flaviviruses detected in mosquitoes in the city of São Paulo.Therefore, mosquitoes previously collected in the city of São Paulo, in part of which the presence of ISFs was previously detected, will be processed using different techniques, including nucleic acid extraction, reverse transcription and long PCR with Pan-Flavi primers capable of detecting the different viral species of flaviviruses. Subsequently, the material will be sent for the preparation of the DNA and NGS library, using the HiSeq 2500 Sequencer equipment (Illumina). The sequences obtained will be analyzed by bioinformatics programs and phylogenetic trees will be reconstructed, according to the viruses detected.Thus, the results obtained may contribute to knowledge about flaviviruses, with regard to distribution, frequency, specificity to a host mosquito, genetic variability and interaction between different viral species that can co-infect the same mosquito, thus generating data useful in the control and prevention of arboviruses caused by flaviviruses.