Emerging approaches to mosquito species identification: an overview with emphasis on nanopore sequencing technology

Abstract Correct identification of mosquito species is crucial for effective vector control, pathogen surveillance, and research and can be achieved through various methods. In this study, we present a review of the most commonly used method for mosquito species identification. We also benchmark the results of DNA sequencing on the MinION nanopore sequencing device against the morphological identification technique, which remains the cornerstone of mosquito species identification. By using 95 samples from the field, we targeted 2 mitochondrial genes (cytochrome oxidase subunit 1 and 3), and successfully distinguished 3 Culex species from the Vishnui subroup (Culex pseudovishnui, Cx. tritaeniorhynchus, and Cx. vishnui) using amplicon-based targeted Oxford Nanopore Technology sequencing, whereas the morphological method proved less effective. Although amplicon-based targeted Oxford Nanopore Technology sequencing is associated with significant costs, it shows great potential for improving accuracy in mosquito species identification, if adequate financial resources are available.