Viruses (Oct 2021)

Metagenomic Identification of Viral Sequences in Laboratory Reagents

  • Ashleigh F. Porter,
  • Joanna Cobbin,
  • Ci-Xiu Li,
  • John-Sebastian Eden,
  • Edward C. Holmes

DOI
https://doi.org/10.3390/v13112122
Journal volume & issue
Vol. 13, no. 11
p. 2122

Abstract

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Metagenomic next-generation sequencing has transformed the discovery and diagnosis of infectious disease, with the power to characterise the complete ‘infectome’ (bacteria, viruses, fungi, parasites) of an individual host organism. However, the identification of novel pathogens has been complicated by widespread microbial contamination in commonly used laboratory reagents. Using total RNA sequencing (“metatranscriptomics”) we documented the presence of contaminant viral sequences in multiple ‘blank’ negative control sequencing libraries that comprise a sterile water and reagent mix. Accordingly, we identified 14 viral sequences in 7 negative control sequencing libraries. As in previous studies, several circular replication-associated protein encoding (CRESS) DNA virus-like sequences were recovered in the blank control libraries, as well as contaminating sequences from the Totiviridae, Tombusviridae and Lentiviridae families of RNA virus. These data suggest that viral contamination of common laboratory reagents is likely commonplace and can comprise a wide variety of viruses.

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