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Nucleic Acids Res. 2015 May 26;43(10):e69. doi: 10.1093/nar/gkv180. Epub 2015 Mar 12.

Accurate read-based metagenome characterization using a hierarchical suite of unique signatures.

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Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA.
Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA


A major challenge in the field of shotgun metagenomics is the accurate identification of organisms present within a microbial community, based on classification of short sequence reads. Though existing microbial community profiling methods have attempted to rapidly classify the millions of reads output from modern sequencers, the combination of incomplete databases, similarity among otherwise divergent genomes, errors and biases in sequencing technologies, and the large volumes of sequencing data required for metagenome sequencing has led to unacceptably high false discovery rates (FDR). Here, we present the application of a novel, gene-independent and signature-based metagenomic taxonomic profiling method with significantly and consistently smaller FDR than any other available method. Our algorithm circumvents false positives using a series of non-redundant signature databases and examines Genomic Origins Through Taxonomic CHAllenge (GOTTCHA). GOTTCHA was tested and validated on 20 synthetic and mock datasets ranging in community composition and complexity, was applied successfully to data generated from spiked environmental and clinical samples, and robustly demonstrates superior performance compared with other available tools.

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