Dengue virus sequencing in Aedes aegypti mosquito saliva. Dengue virus

Like other pathogens with high mutation and replication rates, within-host dengue virus (DENV) populations evolve during infection of their main mosquito vector, Aedes aegypti. To date, however, it is unclear to what extent DENV within-host evolution in the mosquito vector influences the transmitted viral population because of the technical difficulty to sequence DENV genomes in mosquito saliva. Here, we achieved DENV full-genome sequencing by pooling saliva samples collected non-sacrificially from 49 to 163 individual Aedes aegypti mosquitoes previously infected with two DENV-1 genotypes. We compared the transmitted viral populations found in the pooled saliva samples collected in time series with the input viral population present in the infectious blood meal. In one saliva sample that was sequenced at sufficient depth to analyze minority variants, there were detectable but modest changes in DENV variant frequencies. In all pooled saliva samples examined, the full-genome consensus sequence of the input viral population was unchanged. Our findings indicate that within-host evolution in the mosquito vector does not seem to significantly influence the DENV consensus sequence when considering the viral population transmitted from a group of mosquitoes.