Species-resolved profiling of microbial functional genes is important for linking microbial community composition to biological function, but existing functional profiling approaches are not designed to systematically define user-specified genes and resolve their contributing species. We developed Vritra (Versatile gene-guided Reads-identification with Impartial Taxonomic Refinement and Assignment), a framework for constructing gene-specific reference databases for metagenomic and metatranscriptomic data. Vritra expands and refines the UniRef-based sequence space for user-specified target genes using sequence-similarity network connectivity and functional annotations, retains related homologs as decoys to reduce assignment ambiguity, and links the resulting sequences to standardized microbial taxonomy. Across genes involved in oxalate, urate, and bile acid metabolism, Vritra recovered >97% of sequences represented by established annotation resources for most evaluated genes while substantially expanding the represented sequence space. In real microbiome datasets, the expanded references increased recovery of target-gene reads by up to threefold for poorly annotated genes while maintaining high sequence identity. Application to population-based and publicly available microbiome datasets enabled species-resolved profiling of functional genes and revealed gene-specific associations with microbial taxa, dietary factors, and disease-related phenotypes. Vritra provides a scalable framework for translating continuously expanding sequence resources into gene-specific, species-resolved references for microbiome studies.
Li, H., Zhou, B., Liu, M., Curhan, G., Choi, H., Nazzal, L., Peck, g.
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