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Updated: Jun 26, 2026
Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
Published on: May 26, 2013
Evaluation of a diagnostic metagenomic sequencing assay: Virus detection sensitivity and background nucleic acids in
Martin Ekman1, Amir Nematollahi Mahani1, Shambhu Ganeshappa Aralaguppe1
1Department of Clinical Microbiology, Karolinska University Hospital, Stockholm, Sweden.
Background:
Metagenomic sequencing has emerged as an attractive, general, and agnostic diagnostic method, in particular for detection of viruses. However, its application faces limitations, including reduced sensitivity due to background nucleic acid content of samples, and the search for an optimized protocol is still ongoing.
Methods:
We report the development of a metagenomic sequencing protocol for diagnostic use and its performance in detecting DNA and RNA viruses in three different sample materials: serum, cerebrospinal fluid (CSF) and nasopharyngeal swabs (NPS).
Results:
Sensitivity was higher for RNA viruses than for DNA viruses, and also higher in CSF than in serum and lowest in NPS. We characterized the background nucleic acids and found higher DNA than RNA levels in CSF and serum and overall highest nucleic acid levels in NPS, intermediate in serum and lowest in CSF. These differences largely explained the observed variability in sensitivity between sample preparations and sample materials.
Conclusions:
Our results highlight the need to consider sample-type specific characteristics in efforts to improve the sensitivity of metagenomic assays e.g. via host depletion protocols.
