Banca de DEFESA: EVALDO JOAQUIM DE FARIAS FILHO

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : EVALDO JOAQUIM DE FARIAS FILHO
DATE: 16/02/2023
TIME: 14:00
LOCAL: Online
TITLE:

ESTABLISHMENT OF A PROTOCOL FOR METAVIROME ANALYSIS FROM SAMPLES DERIVED FROM MOSQUITOES AND CELL CULTURES.

KEY WORDS:

NGS, vector mosquitoes, DENV, Flaviviridae, Togaviridae e Parvoviridae

PAGES: 72
BIG AREA: Ciências Biológicas
AREA: Genética
SUBÁREA: Genética Molecular e de Microorganismos
SUMMARY:

Viruses identified in mosquitoes are subdivided into two groups: arboviruses, transmitted to vertebrates and able to replicate in both vectors and hosts, and insect-specific viruses (ISVs), which have replication capacity restricted to the invertebrate host. ISVs have the capacity to modulate responses and effects against arboviruses when co-infected in mosquitoes, therefore, the prospection of these ISVs becomes important for analyzes of possible interferences in arbovirology studies. In the last decade, the use of metagenomics tools in the surveillance of medically important arboviruses has been shown to be efficient in the discovery of new ISVs, due to the sensitivity and high throughput of the techniques employed in next-generation sequencing (NGS). Therefore, the objective of this work was to establish a functional protocol for the detection of ISVs in mosquitoes and (C6/36) cells from laboratory strains, and perform the identification of ISVs in the respective groups by NGS. Two methods for virus detection were tested: conventional PCR using degenerate primers for flaviviruses and alphaviruses, using Cx. quinquefasciatus and Ae. aegypti sample and next-generation sequencing, in samples of laboratory mosquitoes and C6/36 cells. It was not possible to detect ISVs using the conventional PCR technique. Metavirome analysis by NGS reveals three viruses: Culex bunyavirus and Wutai mosquito phasivirus in samples of Cx. quinquefasciatus. Culex densovirus was detected in three of four supernatant samples from C6/36 cells co-infected with dengue serotypes 1, 2 and 3. Culex densovirus had 100% coverage of the genome with a depth of 21.872 x. The results bring an efficient metavirome protocol in the detection of ISVs and arboviruses, in addition, this work reinforces the need for metavirome analysis in arbovirology studies.

COMMITTEE MEMBERS:
Presidente - ***.976.374-** - MARCELO HENRIQUE SANTOS PAIVA - UFPE
Interno - JORGE BRAZ TORRES
Interno - JOSE DIJAIR ANTONINO DE SOUZA JUNIOR
Externo à Instituição - DERCILIANO LOPES DA CRUZ - Fiocruz - PE
Externo à Instituição - RENATO ANTÔNIO DOS SANTOS OLIVEIRA - UFPB