Abstract
To improve the storage and transport of clinical specimens for the diagnosis of Neisseria meningitidis (Nm) infections in resource-limited settings, we have evaluated the performance of dried blood spot (DBS) and dried cerebrospinal fluid spot (DCS) assays. DBS and DCS were prepared on filter paper from liquid specimens previously tested for Nm in the United Kingdom. Nm was detected and genogrouped by real-time PCR performed on crude genomic DNA extracted from the DBS (n = 226) and DCS (n = 226) specimens. Targeted whole-genome sequencing was performed on a subset of specimens, DBS (n = 4) and DCS (n = 6). The overall agreement between the analysis of liquid and dried specimens was (94.2%; 95% CI 90.8–96.7) for blood and (96.4%; 95% CI 93.5–98.0) for cerebrospinal fluid. Relative to liquid specimens as the reference, the DBS and DCS assays had sensitivities of (89.1%; 95% CI 82.7–93.8) and (94.2%; 95% CI 88.9–97.5), respectively, and both assays had specificities above 98%. A genogroup was identified by dried specimen analysis for 81.9% of the confirmed meningococcal infections. Near full-length Nm genome sequences (>86%) were obtained for all ten specimens tested which allowed determination of the sequence type, clonal complex, presence of antimicrobial resistance and other meningococcal genotyping. Dried blood and CSF filter spot assays offer a practical alternative to liquid specimens for the molecular and genomic characterisation of invasive meningococcal diseases in low-resource settings.
Original language | English |
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Article number | 11879 |
Journal | International Journal of Molecular Sciences |
Volume | 23 |
Issue number | 19 |
DOIs | |
Publication status | Published - Oct 2022 |
Bibliographical note
Funding Information:This work was funded in part through a Global Challenges Research Funds (GCRF): UCL Internal Small Grant to B.K.-A. B.K.-A., N.T., and R.H. were supported by the NIHR Global Health Research Unit on Mucosal Pathogens using UK aid from the UK Government. RSH is an NIHR Senior Investigator. The UK Health Security Agency provided staff and facility costs for work performed at the Meningococcal Reference Unit. The views expressed in this publication are those of the author(s) and not necessarily those of the NIHR, the Department of Health and Social Care, or the authors’ affiliated institutions.
Publisher Copyright:
© 2022 by the authors.
Keywords
- Neisseria meningitidis
- bacteraemia
- blood
- cerebrospinal fluid
- diagnosis
- dried filter paper spot
- invasive meningococcal disease
- meningitis
- real-time PCR
- targeted enrichment
- whole genome sequencing