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bioRxiv · 10.1101/2020.04.25.060947

MINERVA: A facile strategy for SARS-CoV-2 whole genome deep sequencing of clinical samples

Chen, C.·Li, J.·Di, L.·Jing, Q.·Du, P.·Song, C.·Li, Q.·Cao, Y.·Xie, S.·Wu, A. R.·Zeng, H.·Huang, Y.·Wang, J.
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Abstract

The novel coronavirus disease 2019 (COVID-19) pandemic poses a serious public health risk. Analyzing the genome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from clinical samples is crucial for the understanding of viral spread and viral evolution, as well as for vaccine development. Existing sample preparation methods for viral genome sequencing are demanding on user technique and time, and thus not ideal for time-sensitive clinical samples; these methods are also not optimized for high performance on viral genomes. We have developed MetagenomIc RNA EnRichment VirAl sequencing (MINERVA), a facile, practical, and robust approach for metagenomic and deep viral sequencing from clinical samples. This approach uses direct tagmentation of RNA/DNA hybrids using Tn5 transposase to greatly simplify the sequencing library construction process, while subsequent targeted enrichment can generate viral genomes with high sensitivity, coverage, and depth. We demonstrate the utility of MINERVA on pharyngeal, sputum and stool samples collected from COVID-19 patients, successfully obtaining both whole metatranscriptomes and complete high-depth high-coverage SARS-CoV-2 genomes from these clinical samples, with high yield and robustness. MINERVA is compatible with clinical nucleic extracts containing carrier RNA. With a shortened hands-on time from sample to virus-enriched sequencing-ready library, this rapid, versatile, and clinic-friendly approach will facilitate monitoring of viral genetic variations during outbreaks, both current and future.

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Chen, C., Li, J., Di, L., Jing, Q., Du, P., Song, C., Li, Q., Cao, Y., Xie, S., Wu, A. R., Zeng, H., Huang, Y., Wang, J.. 2020-04-25. MINERVA: A facile strategy for SARS-CoV-2 whole genome deep sequencing of clinical samples. https://doi.org/10.1101/2020.04.25.060947

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