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Volume 10, Issue 1

Tracking arboviruses, their transmission vectors and potential hosts by nanopore sequencing of mosquitoes Open Access

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 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.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): arboviruses , cytochrome c oxidase , metagenomics , mosquitoes , nanopore and SMART-9N
Funding
This study was supported by the:
  • CNPq (National Council for Scientific and Technological Development) (Award 309396/2021-2)
    • Principle Award Recipient: KarinKirchgatter
  • Wellcome ARTIC network (Award 206298)
    • Principle Award Recipient: NicholasJ. Loman
  • Wellcome ARTIC network (Award 206298)
    • Principle Award Recipient: JoshuaQuick
  • Medical Research Council-São Paulo Research Foundation (FAPESP) CADDE partnership award (Award MRC MR/S0195/1 and FAPESP 18/14389-0)
    • Principle Award Recipient: NunoR. Faria
  • Bill and Melinda Gates Foundation (Award INV-034540)
    • Principle Award Recipient: IngraM. Claro
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2018/16232-1)
    • Principle Award Recipient: Liliande Oliveira Guimarães
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2017/50345-5)
    • Principle Award Recipient: JeremyDarius Mirza
© 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.
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2024-01-19
2024-05-09
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Tracking arboviruses, their transmission vectors and potential hosts by nanopore sequencing of mosquitoes
M Gen 10, 001184 (2024); https://doi.org/10.1099/mgen.0.001184
/content/journal/mgen/10.1099/mgen.0.001184
/content/journal/mgen/10.1099/mgen.0.001184
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