COMPARISON OF NANOPORE AND CLASSICAL SANGER SEQUENCING TO IDENTIFY MOSQUITO BLOODMEAL HOSTS.

Abstract

The tools available to vector control districts (VCDs) to collect mosquito surveillance data are constantly evolving. As more VCDs obtain real-time polymerase chain reaction (PCR) instruments and the costs associated with computing power and next-generation sequencing continue to decrease, the option of generating useful molecular data in-house becomes more viable. Measures such as arbovirus testing and genotyping for insecticide resistance mutations using RT-qPCR, and identifying species used for mosquito bloodmeals with next-generation sequencing or Sanger sequencing are examples. In this study we identify mosquito host bloodmeal species using Nanopore sequencing from Oxford Nanopore Technologies. We used MinION and Flongle flow cells and a Mk1C device to sequence 96 barcoded amplicon samples in a single sequencing run, and share details of data analysis using the free-to-use Galaxy bioinformatics platform. After sequencing the same samples with Sanger sequencing, we conclude that Nanopore sequencing is better at identifying species in mixed bloodmeals. This work demonstrates a potential use of nanopore sequencing by VCDs with basic biology laboratory and computing equipment.