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Bacterial ribosomal RNA detection in cerebrospinal fluid using a viromics approach

Abstract:
Abstract Background In patients with central nervous system (CNS) infections identification of the causative pathogen is important for treatment. Metagenomic next-generation sequencing techniques are increasingly being applied to identify causes of CNS infections, as they can detect any pathogen nucleic acid sequences present. Viromic techniques that enrich samples for virus particles prior to sequencing may simultaneously enrich ribosomes from bacterial pathogens, which are similar in size to small viruses. Methods We studied the performance of a viromic library preparation technique (VIDISCA) combined with low-depth IonTorrent sequencing (median ~ 25,000 reads per sample) for detection of ribosomal RNA from common pathogens, analyzing 89 cerebrospinal fluid samples from patients with culture proven bacterial meningitis. Results Sensitivity and specificity to Streptococcus pneumoniae (n = 24) before and after optimizing threshold parameters were 79% and 52%, then 88% and 90%. Corresponding values for Neisseria meningitidis (n = 22) were 73% and 93%, then 67% and 100%, Listeria monocytogenes (n = 24) 21% and 100%, then 27% and 100%, and Haemophilus influenzae (n = 18) 56% and 100%, then 71% and 100%. A higher total sequencing depth, no antibiotic treatment prior to lumbar puncture, increased disease severity, and higher c-reactive protein levels were associated with pathogen detection. Conclusion We provide proof of principle that a viromic approach can be used to correctly identify bacterial ribosomal RNA in patients with bacterial meningitis. Further work should focus on increasing assay sensitivity, especially for problematic species (e.g. L. monocytogenes), as well as profiling additional pathogens. The technique is most suited to research settings and examination of idiopathic cases, rather than an acute clinical setting.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1186/s12987-022-00400-5

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Institution:
University of Oxford
Role:
Author
ORCID:
0000-0001-9865-4608
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Role:
Author
ORCID:
0000-0001-9264-5874
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Role:
Author
ORCID:
0000-0002-1085-2093
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Role:
Author
ORCID:
0000-0002-4571-044X


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Funder identifier:
10.13039/501100001826
Grant:
917.17.308
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Funder identifier:
10.13039/501100000781
Grant:
101001237


Publisher:
BioMed Central
Journal:
Fluids and Barriers of the CNS More from this journal
Volume:
19
Issue:
1
Pages:
102-102
Article number:
102
Publication date:
2022-12-22
DOI:
EISSN:
2045-8118
ISSN:
2045-8118


Language:
English
Keywords:
Pubs id:
1347940
Local pid:
pubs:1347940
Source identifiers:
W4312190515
Deposit date:
2026-05-08
ARK identifier:
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