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Benchmark of thirteen bioinformatic pipelines for metagenomic virus diagnostics using datasets from clinical samples.

ΤίτλοςBenchmark of thirteen bioinformatic pipelines for metagenomic virus diagnostics using datasets from clinical samples.
Publication TypeJournal Article
Year of Publication2021
Authorsde Vries, J. J. C., Brown J. R., Fischer N., Sidorov I. A., Morfopoulou S., Huang J., Munnink B. B. Oude, Sayiner A., Bulgurcu A., Rodriguez C., Gricourt G., Keyaerts E., Beller L., Bachofen C., Kubacki J., Samuel C., Florian L., Dennis S., Beer M., Hoeper D., Huber M., Kufner V., Zaheri M., Lebrand A., Papa A., van Boheemen S., Kroes A. C. M., Breuer J., F Lopez-Labrador X., & Claas E. C. J.
JournalJ Clin Virol
Volume141
Pagination104908
Date Published2021 Aug
ISSN1873-5967
Abstract

INTRODUCTION: Metagenomic sequencing is increasingly being used in clinical settings for difficult to diagnose cases. The performance of viral metagenomic protocols relies to a large extent on the bioinformatic analysis. In this study, the European Society for Clinical Virology (ESCV) Network on NGS (ENNGS) initiated a benchmark of metagenomic pipelines currently used in clinical virological laboratories.
METHODS: Metagenomic datasets from 13 clinical samples from patients with encephalitis or viral respiratory infections characterized by PCR were selected. The datasets were analyzed with 13 different pipelines currently used in virological diagnostic laboratories of participating ENNGS members. The pipelines and classification tools were: Centrifuge, DAMIAN, DIAMOND, DNASTAR, FEVIR, Genome Detective, Jovian, MetaMIC, MetaMix, One Codex, RIEMS, VirMet, and Taxonomer. Performance, characteristics, clinical use, and user-friendliness of these pipelines were analyzed.
RESULTS: Overall, viral pathogens with high loads were detected by all the evaluated metagenomic pipelines. In contrast, lower abundance pathogens and mixed infections were only detected by 3/13 pipelines, namely DNASTAR, FEVIR, and MetaMix. Overall sensitivity ranged from 80% (10/13) to 100% (13/13 datasets). Overall positive predictive value ranged from 71-100%. The majority of the pipelines classified sequences based on nucleotide similarity (8/13), only a minority used amino acid similarity, and 6 of the 13 pipelines assembled sequences de novo. No clear differences in performance were detected that correlated with these classification approaches. Read counts of target viruses varied between the pipelines over a range of 2-3 log, indicating differences in limit of detection.
CONCLUSION: A wide variety of viral metagenomic pipelines is currently used in the participating clinical diagnostic laboratories. Detection of low abundant viral pathogens and mixed infections remains a challenge, implicating the need for standardization and validation of metagenomic analysis for clinical diagnostic use. Future studies should address the selective effects due to the choice of different reference viral databases.

DOI10.1016/j.jcv.2021.104908
Alternate JournalJ Clin Virol
PubMed ID34273858

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