ViroSeek: a viral detection pipeline for second-generation sequencing

Abstract Arbovirus emergences represent a rising public health issue and are exacerbated by climate change and globalization. Virome analysis has become a key approach for monitoring and managing infectious diseases, yet existing tools often remain technically complex and inaccessible to non-specialists. In this context, we present ViroSeek, a lightweight, reproducible and accessible bioinformatics pipeline specifically designed for the taxonomic analysis of second-generation sequencing data. ViroSeek performs a series of automated steps: quality control (FastQC), trimming (TrimGalore or fastp), host and bacterial sequence removal (BBduk), assembly (SPAdes), taxonomic assignment (DIAMOND and TaxonKit), read remapping for quantification (minimap2), and PCR duplicate removal (Samtools). The whole process is designed to produce a clear, usable viral taxonomy table that is suitable for diversity studies. ViroSeek was empirically validated on enriched control samples containing a known panel of viruses. All the expected viruses were correctly detected. Bacterial and host contaminant sequences were effectively removed. The pipeline is freely available and fully documented, supporting its adoption and adaptation by the research community. It provides an optimized solution for virome analysis.