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OptiFit: an improved method for fitting amplicon sequences to existing OTUs

View ORCID ProfileKelly L. Sovacool, Sarah L. Westcott, M. Brodie Mumphrey, View ORCID ProfileGabrielle A. Dotson, View ORCID ProfilePatrick D. Schloss
doi: https://doi.org/10.1101/2021.11.09.468000
Kelly L. Sovacool
1Department of Computational Medicine and Bioinformatics, University of Michigan
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Sarah L. Westcott
2Department of Microbiology and Immunology, University of Michigan
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M. Brodie Mumphrey
1Department of Computational Medicine and Bioinformatics, University of Michigan
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Gabrielle A. Dotson
1Department of Computational Medicine and Bioinformatics, University of Michigan
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Patrick D. Schloss
2Department of Microbiology and Immunology, University of Michigan
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  • For correspondence: pschloss@umich.edu
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Abstract

Assigning amplicon sequences to operational taxonomic units (OTUs) is often an important step in characterizing the composition of microbial communities across large datasets. OptiClust, a de novo OTU clustering method, has been shown to produce higher quality OTU assignments than other methods and at comparable or faster speeds. A notable difference between de novo clustering and database-dependent reference clustering methods is that OTU assignments from de novo methods may change when new sequences are added to a dataset. However, in some cases one may wish to incorporate new samples into a previously clustered dataset without performing clustering again on all sequences, such as when comparing across datasets or deploying machine learning models where OTUs are features. Existing reference-based clustering methods produce consistent OTUs, but they only consider the similarity of each query sequence to a single reference sequence in an OTU, thus resulting in OTU assignments that are significantly worse than those generated by de novo methods. To provide an efficient and robust method to fit amplicon sequence data to existing OTUs, we developed the OptiFit algorithm. Inspired by OptiClust, OptiFit considers the similarity of all pairs of reference and query sequences in an OTU to produce OTUs of the best possible quality. We tested OptiFit using four microbiome datasets with two different strategies: by clustering to an external reference database or by splitting the dataset into a reference and query set and clustering the query sequences to the reference set after clustering it using OptiClust. The result is an improved implementation of closed and open-reference clustering. OptiFit produces OTUs of similar quality as OptiClust and at faster speeds when using the split dataset strategy, although the OTU quality and processing speed depends on the database chosen when using the external database strategy. OptiFit provides a suitable option for users who require consistent OTU assignments at the same quality afforded by de novo clustering methods.

Importance Advancements in DNA sequencing technology have allowed researchers to affordably generate millions of sequence reads from microorganisms in diverse environments. Efficient and robust software tools are needed to assign microbial sequences into taxonomic groups for characterization and comparison of communities. The OptiClust algorithm produces high quality groups by comparing sequences to each other, but the assignments can change when new sequences are added to a dataset, making it difficult to compare different studies. Other approaches assign sequences to groups by comparing them to sequences in a reference database to produce consistent assignments, but the quality of the groups produced is reduced compared to OptiClust. We developed OptiFit, a new reference-based algorithm that produces consistent yet high quality assignments like OptiClust. OptiFit allows researchers to compare microbial communities across different studies or add new data to existing studies without sacrificing the quality of the group assignments.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted November 11, 2021.
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OptiFit: an improved method for fitting amplicon sequences to existing OTUs
Kelly L. Sovacool, Sarah L. Westcott, M. Brodie Mumphrey, Gabrielle A. Dotson, Patrick D. Schloss
bioRxiv 2021.11.09.468000; doi: https://doi.org/10.1101/2021.11.09.468000
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OptiFit: an improved method for fitting amplicon sequences to existing OTUs
Kelly L. Sovacool, Sarah L. Westcott, M. Brodie Mumphrey, Gabrielle A. Dotson, Patrick D. Schloss
bioRxiv 2021.11.09.468000; doi: https://doi.org/10.1101/2021.11.09.468000

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