Amplicon sequencing is an effective and increasingly applied method for
studying viral communities in the environment. Here, we present
vAMPirus, a user-friendly, comprehensive, and versatile DNA and RNA
virus amplicon sequence analysis program, designed to support
investigators in exploring virus amplicon sequencing data and running
informed, reproducible analyses. vAMPirus intakes raw virus amplicon
libraries and, by default, performs nucleotide- and protein-based
analyses to produce results such as sequence abundance information,
taxonomic classifications, phylogenies, and community diversity metrics.
The vAMPirus pipelines additionally include optional approaches that can
increase the biological signal-to-noise ratio in results by leveraging
tools not yet commonly applied to virus amplicon data analyses. In this
paper, we validate the vAMPirus analytical framework and illustrate its
implementation into the general virus amplicon sequencing workflow by
recapitulating findings from two previously published double-stranded
DNA virus datasets. As a case study, we also apply the program to
explore the diversity and distribution of a coral reef-associated RNA
virus. vAMPirus is incorporated with the Nextflow workflow manager,
offering straightforward scalability, standardization, and communication
of virus lineage-specific analyses. The vAMPirus framework itself is
also designed to be adaptable; community-driven analytical standards
will continue to be incorporated as the field advances. vAMPirus
supports researchers in revealing patterns of virus diversity and
population dynamics in nature, while promoting study reproducibility and
comparability.