Tracking arboviruses, their transmission vectors and potential hosts by nanopore sequencing of mosquitoes
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
The risk to human health from mosquito-borne viruses such as dengue, chikungunya and yellow fever is increasing due to increased human expansion, deforestation and climate change. To anticipate and predict the spread and transmission of mosquito-borne viruses, a better understanding of the transmission cycle in mosquito populations is needed. We present a pathogen-agnostic combined sequencing protocol for identifying vectors, viral pathogens and their hosts or reservoirs using portable Oxford Nanopore sequencing. Using mosquitoes collected in São Paulo, Brazil, we extracted RNA for virus identification and DNA for blood meal and mosquito identification. Mosquitoes and blood meals were identified by comparing cytochrome c oxidase I (COI) sequences against a curated Barcode of Life Data System (BOLD). Viruses were identified using the SMART-9N protocol, which allows amplified DNA to be prepared with native barcoding for nanopore sequencing. Kraken 2 was employed to detect viral pathogens and Minimap2 and BOLD identified the contents of the blood meal. Due to the high similarity of some species, mosquito identification was conducted using blast after generation of consensus COI sequences using RACON polishing. This protocol can simultaneously uncover viral diversity, mosquito species and mosquito feeding habits. It also has the potential to increase understanding of mosquito genetic diversity and transmission dynamics of zoonotic mosquito-borne viruses.
Date Issued
2024-01-01
Date Acceptance
2024-01-08
Citation
Microbial genomics, 2024, 10 (1)
ISSN
2057-5858
Publisher
Microbiology Society
Journal / Book Title
Microbial genomics
Volume
10
Issue
1
Copyright Statement
© 2024 The Authors This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/38240642
Subjects
9N.
arboviruses
BARCODE
BLOOD
CULEX DIPTERA-CULICIDAE
cytochrome c oxidase
Genetics & Heredity
IDENTIFICATION
Life Sciences & Biomedicine
metagenomics
Microbiology
mosquitoes
nanopore
Science & Technology
SMART
ZIKA VIRUS
Publication Status
Published
Coverage Spatial
England
Article Number
001184
Date Publish Online
2024-01-19