{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T03:19:11Z","timestamp":1788923951823,"version":"build-2803163510"},"reference-count":37,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2022,10,6]],"date-time":"2022-10-06T00:00:00Z","timestamp":1665014400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Global Challenges Research Funds (GCRF): UCL Internal Small Grant"},{"name":"NIHR Global Health Research Unit on Mucosal Pathogens"},{"name":"UK Government"},{"name":"UK Health Security Agency"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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\u201396.7) for blood and (96.4%; 95% CI 93.5\u201398.0) for cerebrospinal fluid. Relative to liquid specimens as the reference, the DBS and DCS assays had sensitivities of (89.1%; 95% CI 82.7\u201393.8) and (94.2%; 95% CI 88.9\u201397.5), 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 (&gt;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.<\/jats:p>","DOI":"10.3390\/ijms231911879","type":"journal-article","created":{"date-parts":[[2022,10,8]],"date-time":"2022-10-08T23:39:31Z","timestamp":1665272371000},"page":"11879","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1202-8540","authenticated-orcid":false,"given":"Brenda A.","family":"Kwambana-Adams","sequence":"first","affiliation":[{"name":"NIHR Global Health Research Unit on Mucosal Pathogens, Division of Infection and Immunity, University College London, London WC1E 6BT, UK"},{"name":"Clinical Sciences Department, Liverpool School of Tropical Medicine, Liverpool L3 5QA, UK"},{"name":"Malawi-Liverpool-Wellcome Clinical Research Programme (MLW), Blantyre P.O. Box 30096, Malawi"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4916-3623","authenticated-orcid":false,"given":"Stephen A.","family":"Clark","sequence":"additional","affiliation":[{"name":"Meningococcal Reference Unit, United Kingdom Health Security Agency (UKHSA), Manchester M13 9WL, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6294-5630","authenticated-orcid":false,"given":"Nicole","family":"Tay","sequence":"additional","affiliation":[{"name":"NIHR Global Health Research Unit on Mucosal Pathogens, Division of Infection and Immunity, University College London, London WC1E 6BT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Schadrac","family":"Agbla","sequence":"additional","affiliation":[{"name":"Department of Health Data Science, University of Liverpool, Liverpool L69 3GF, UK"},{"name":"Department of Infectious Disease Epidemiology, London School of Hygiene and Tropical Medicine, London WC1E 7HT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chrispin","family":"Chaguza","sequence":"additional","affiliation":[{"name":"NIHR Global Health Research Unit on Mucosal Pathogens, Division of Infection and Immunity, University College London, London WC1E 6BT, UK"},{"name":"Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK"},{"name":"Department of Epidemiology of Microbial Diseases, Yale School of Public Health, Yale University, New Haven, CT 06510, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eunice W.","family":"Kagucia","sequence":"additional","affiliation":[{"name":"Department of Epidemiology and Demography, KEMRI-Wellcome Trust Research Programme, Kilifi P.O. Box 230-8010, Kenya"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ray","family":"Borrow","sequence":"additional","affiliation":[{"name":"Meningococcal Reference Unit, United Kingdom Health Security Agency (UKHSA), Manchester M13 9WL, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4573-449X","authenticated-orcid":false,"given":"Robert S.","family":"Heyderman","sequence":"additional","affiliation":[{"name":"NIHR Global Health Research Unit on Mucosal Pathogens, Division of Infection and Immunity, University College London, London WC1E 6BT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"B26","DOI":"10.1016\/j.vaccine.2011.12.032","article-title":"The changing and dynamic epidemiology of meningococcal disease","volume":"30","author":"Halperin","year":"2012","journal-title":"Vaccine"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1007\/978-1-61779-346-2_2","article-title":"Classification and pathogenesis of meningococcal infections","volume":"799","author":"Brandtzaeg","year":"2012","journal-title":"Methods Mol. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"867","DOI":"10.1016\/S1473-3099(17)30301-8","article-title":"Impact of MenAfriVac in nine countries of the African meningitis belt, 2010-15: An analysis of surveillance data","volume":"17","author":"Trotter","year":"2017","journal-title":"Lancet Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"14194","DOI":"10.1038\/s41598-018-32475-2","article-title":"Meningococcus serogroup C clonal complex ST-10217 outbreak in Zamfara State, Northern Nigeria","volume":"8","author":"Amaza","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1288","DOI":"10.1016\/S1473-3099(16)30253-5","article-title":"Emergence of epidemic Neisseria meningitidis serogroup C in Niger, 2015: An analysis of national surveillance data","volume":"16","author":"Sidikou","year":"2016","journal-title":"Lancet Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Retchless, A.C., Congo-Ouedraogo, M., Kambire, D., Vuong, J., Chen, A., Hu, F., Ba, A.K., Ouedraogo, A.S., Hema-Ouangraoua, S., and Patel, J.C. (2018). Molecular characterization of invasive meningococcal isolates in Burkina Faso as the relative importance of serogroups X and W increases, 2008\u20132012. BMC Infect. Dis., 18.","DOI":"10.1186\/s12879-018-3247-x"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5510","DOI":"10.1073\/pnas.1802298115","article-title":"Acquisition of virulence genes by a carrier strain gave rise to the ongoing epidemics of meningococcal disease in West Africa","volume":"115","author":"Brynildsrud","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Tzeng, Y.L., and Stephens, D.S. (2021). A Narrative Review of the W, X, Y, E, and NG of Meningococcal Disease: Emerging Capsular Groups, Pathotypes, and Global Control. Microorganisms, 9.","DOI":"10.3390\/microorganisms9030519"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1088","DOI":"10.1016\/S1473-3099(18)30400-6","article-title":"Safety and immunogenicity of a pentavalent meningococcal conjugate vaccine containing serogroups A, C, Y, W, and X in healthy adults: A phase 1, single-centre, double-blind, randomised, controlled study","volume":"18","author":"Chen","year":"2018","journal-title":"Lancet. Infect. Dis."},{"key":"ref_10","unstructured":"World Health Organization (2015). Preparedness for outbreaks of meningococcal meningitis due to Neisseria meningitidis serogroup C in Africa: Recommendations from a WHO expert consultation. Relev. Epidemiol. Hebd., 90, 633\u2013636."},{"key":"ref_11","unstructured":"WHO (2021, November 26). Defeating Meningitis by 2030: A Global Road Map, Available online: https:\/\/www.who.int\/publications\/i\/item\/9789240026407."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Stresman, G.H., Mwesigwa, J., Achan, J., Giorgi, E., Worwui, A., Jawara, M., Di Tanna, G.L., Bousema, T., Van Geertruyden, J.P., and Drakeley, C. (2018). Do hotspots fuel malaria transmission: A village-scale spatio-temporal analysis of a 2-year cohort study in The Gambia. BMC Med., 16.","DOI":"10.1186\/s12916-018-1141-4"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1136\/sextrans-2018-053567","article-title":"Dried blood spot and mini-tube blood sample collection kits for postal HIV testing services: A comparative review of successes in a real-world setting","volume":"95","author":"Page","year":"2018","journal-title":"Sex. Transm. Infect."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Iroh Tam, P.Y., Hernandez-Alvarado, N., Schleiss, M.R., Yi, A.J., Hassan-Hanga, F., Onuchukwu, C., Umoru, D., and Obaro, S.K. (2018). Detection of Streptococcus pneumoniae from culture-negative dried blood spots by real-time PCR in Nigerian children with acute febrile illness. BMC Res. Notes, 11.","DOI":"10.1186\/s13104-018-3770-2"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Pimenta, F.C., Moiane, B., Lessa, F.C., Venero, A.L., Moura, I., Larson, S., Massora, S., Chauque, A., Tembe, N., and Mucavele, H. (2020). Dried blood spots for Streptococcus pneumoniae and Haemophilus influenzae detection and serotyping among children < 5 years old in rural Mozambique. BMC Pediatr., 20.","DOI":"10.1186\/s12887-020-02209-3"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/0035-9203(76)90042-0","article-title":"The bacteriological diagnosis of pyogenic meningitis from cerebrospinal fluid dried on filter paper","volume":"70","author":"Whittle","year":"1976","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Cogley, M.F., Wiberley-Bradford, A.E., Mochal, S.T., Dawe, S.J., Piro, Z.D., and Baker, M.W. (2021). Newborn Screening for Severe Combined Immunodeficiency Using the Multiple of the Median Values of T-Cell Receptor Excision Circles. Int. J. Neonatal Screen., 7.","DOI":"10.3390\/ijns7030043"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"S49","DOI":"10.1093\/cid\/ciz472","article-title":"Pediatric Bacterial Meningitis Surveillance in the World Health Organization African Region Using the Invasive Bacterial Vaccine-Preventable Disease Surveillance Network, 2011\u20132016","volume":"69","author":"Mwenda","year":"2019","journal-title":"Clin. Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Vuong, J., Collard, J.M., Whaley, M.J., Bassira, I., Seidou, I., Diarra, S., Ouedraogo, R.T., Kambire, D., Taylor, T.H., and Sacchi, C. (2016). Development of Real-Time PCR Methods for the Detection of Bacterial Meningitis Pathogens without DNA Extraction. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0147765"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1507","DOI":"10.3201\/eid1909.130077","article-title":"Serogroup W135 meningococcal disease, The Gambia, 2012","volume":"19","author":"Hossain","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Fukasawa, L.O., Goncalves, M.G., Higa, F.T., Castilho, E.A., Ibarz-Pavon, A.B., and Sacchi, C.T. (2017). Use of cerebrospinal fluid and serum samples impregnated on FTATM Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0172794"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1093\/trstmh\/trab107","article-title":"Real-time polymerase chain reaction on filter paper spotted samples: A gateway to molecular diagnosis of invasive bacterial diseases for rural areas in low-income countries","volume":"116","author":"Ricci","year":"2022","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Iroh Tam, P.Y., Hernandez-Alvarado, N., Schleiss, M.R., Hassan-Hanga, F., Onuchukwu, C., Umoru, D., and Obaro, S.K. (2016). Molecular Detection of Streptococcus pneumoniae on Dried Blood Spots from Febrile Nigerian Children Compared to Culture. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0152253"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2009","DOI":"10.1007\/s00216-020-02490-2","article-title":"PCR inhibition in qPCR, dPCR and MPS-mechanisms and solutions","volume":"412","author":"Sidstedt","year":"2020","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2569","DOI":"10.1007\/s00216-018-0931-z","article-title":"Inhibition mechanisms of hemoglobin, immunoglobulin G, and whole blood in digital and real-time PCR","volume":"410","author":"Sidstedt","year":"2018","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e01780-20","DOI":"10.1128\/JCM.01780-20","article-title":"Full Molecular Typing of Neisseria meningitidis Directly from Clinical Specimens for Outbreak Investigation","volume":"58","author":"Itsko","year":"2020","journal-title":"J. Clin. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1093\/infdis\/jiac104","article-title":"Enhancing Meningococcal Genomic Surveillance in the Meningitis Belt Using High-Resolution Culture-Free Whole-Genome Sequencing","volume":"226","author":"Itsko","year":"2022","journal-title":"J. Infect. Dis."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1016\/j.jinf.2013.06.007","article-title":"Added value of PCR-testing for confirmation of invasive meningococcal disease in England","volume":"67","author":"Heinsbroek","year":"2013","journal-title":"J. Infect."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1099\/jmm.0.46288-0","article-title":"Epidemiology of meningococcal disease in England and Wales 1993\/94 to 2003\/04: Contribution and experiences of the Meningococcal Reference Unit","volume":"55","author":"Gray","year":"2006","journal-title":"J. Med. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.diagmicrobio.2004.04.013","article-title":"Comparison of four polymerase chain reaction assays for specificity in the identification of Streptococcus pneumoniae","volume":"49","author":"Messmer","year":"2004","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_31","unstructured":"Dolan, J.M., Wang, X., Theodore, J., Hatcher, C., Carvalho, M.G., Harcourt, B., Pimenta, F.C., Beall, B., Edmond, K., and Mendisaihan, J. (2008, January 7\u201312). Improved Real-time PCR to Detect N. meningitidis, S. pneumoniae and H. influenzae from international clinical specimens. Proceedings of the 16th International Pathogenic Neisseria Conference (IPNC), Rotterdam, The Netherlands."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4546","DOI":"10.1128\/iai.61.11.4546-4552.1993","article-title":"Limited diversity of the protein D gene (hpd) among encapsulated and nonencapsulated Haemophilus influenzae strains","volume":"61","author":"Janson","year":"1993","journal-title":"Infect. Immun."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/j.ijmm.2017.11.004","article-title":"Targeted DNA enrichment and whole genome sequencing of Neisseria meningitidis directly from clinical specimens","volume":"308","author":"Clark","year":"2018","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"10","DOI":"10.14806\/ej.17.1.200","article-title":"Cutadapt removes adapter sequences from high-throughput sequencing reads","volume":"17","author":"Martin","year":"2011","journal-title":"EMBnet. J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1093\/bioinformatics\/btr026","article-title":"Quality control and preprocessing of metagenomic datasets","volume":"27","author":"Schmieder","year":"2011","journal-title":"Bioinformatics"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1072","DOI":"10.1093\/bioinformatics\/btt086","article-title":"QUAST: Quality assessment tool for genome assemblies","volume":"29","author":"Gurevich","year":"2013","journal-title":"Bioinformatics"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Jolley, K.A., and Maiden, M.C. (2010). BIGSdb: Scalable analysis of bacterial genome variation at the population level. BMC Bioinform., 11.","DOI":"10.1186\/1471-2105-11-595"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/19\/11879\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:47:25Z","timestamp":1760143645000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/19\/11879"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,6]]},"references-count":37,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["ijms231911879"],"URL":"https:\/\/doi.org\/10.3390\/ijms231911879","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,6]]}}}