Evaluation of a diagnostic metagenomic sequencing assay: Virus detection sensitivity and background nucleic acids in three different sample materials.
JSON
Ekman M, Mahani AN, Aralaguppe SG, Normark T, Stamouli S, Andersson-Li L, Sun D, Broddesson S, Wirta V, Björkström NK, Albert J, Allander T
J. Clin. Virol. 181 (-) 105882 [2025-10-20; online 2025-10-20]
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. 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). 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. 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.
Clinical Genomics Stockholm [Service]
PubMed 41161015
DOI 10.1016/j.jcv.2025.105882
Crossref 10.1016/j.jcv.2025.105882
pii: S1386-6532(25)00124-6