{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T02:53:19Z","timestamp":1760151199985,"version":"build-2065373602"},"reference-count":41,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,10]],"date-time":"2022-02-10T00:00:00Z","timestamp":1644451200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006478","name":"Federal Ministry of Defence","doi-asserted-by":"publisher","award":["36K2-S-45 1922"],"award-info":[{"award-number":["36K2-S-45 1922"]}],"id":[{"id":"10.13039\/501100006478","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The suitability of incubated blood culture material for forensic molecular malaria diagnosis was assessed for non-endemic settings for cases in which the differential diagnosis malaria was initially overlooked. For the proof-of-principle assessment, residual blood culture materials from febrile patients from tropical Ghana were investigated by real-time PCR and compared with available historic microscopic results. In 2114 samples, for which microscopical results and real-time PCR results were available, microscopical results comprised 711 P. falciparum detections, 7 P. malariae detections, 1 microscopically not-further-discriminable Plasmodium spp. detection as well as 13 detections of mixed infections comprising 12 cases of P. falciparum\/P. malariae co-infections and 1 case of a P. falciparum\/P. ovale complex co-infection, while real-PCR indicated 558 P. falciparum detections, 95 P. malariae detections, 10 P. ovale complex detections, 1 P. vivax detection and 4 detected P. falciparum\/P. malariae co-infections. Concordance of routine microscopy and real-time PCR was imperfect. Using routine microscopy as reference was associated with a seemingly low agreement of positive real-time PCR results of 90.9%. However, if positive samples, either by routine microscopy or real-time PCR or both, were applied as a combined reference, the agreement of positive results obtained with real-time PCR was increased from 74.0% to 77.9%, while the agreement of positive results obtained with routine microscopy was decreased from 100% to 85.3%. The predictive value of routine microscopy for negative results in the reference was slightly better with 90.9% compared to real-time PCR with 86.9%; the concordance between routine microscopy and real-time PCR was imperfect. In conclusion, even suboptimal sample materials such as incubated blood culture materials can be applied for forensic malaria diagnosis, if more suitable sample materials are not available, but the molecular detection rate of positive results in routine microscopy is much lower than previously reported for non-incubated blood.<\/jats:p>","DOI":"10.3390\/microorganisms10020406","type":"journal-article","created":{"date-parts":[[2022,2,11]],"date-time":"2022-02-11T02:35:51Z","timestamp":1644546951000},"page":"406","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Limited Reliability of the Molecular Detection of Plasmodium spp. from Incubated Blood Culture Samples for Forensic Purposes"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9173-7072","authenticated-orcid":false,"given":"Felix","family":"Weinreich","sequence":"first","affiliation":[{"name":"Department of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 20359 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4875-1519","authenticated-orcid":false,"given":"Ralf Matthias","family":"Hagen","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Hospital Hygiene, Bundeswehr Central Hospital Koblenz, 56070 Koblenz, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4261-365X","authenticated-orcid":false,"given":"Wibke","family":"Loag","sequence":"additional","affiliation":[{"name":"Bernhard Nocht Institute for Tropical Medicine Hamburg, 20259 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oumou","family":"Ma\u00efga-Ascofar\u00e9","sequence":"additional","affiliation":[{"name":"Bernhard Nocht Institute for Tropical Medicine Hamburg, 20259 Hamburg, Germany"},{"name":"Kumasi Centre for Collaborative Research in Tropical Medicine, College of Health Sciences, KNUST, Kumasi 039-5028, Ghana"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5432-3217","authenticated-orcid":false,"given":"Denise","family":"Dekker","sequence":"additional","affiliation":[{"name":"Bernhard Nocht Institute for Tropical Medicine Hamburg, 20259 Hamburg, Germany"},{"name":"German Center for Infection Research (DZIF), Partner Site Hamburg-L\u00fcbeck-Borstel-Riems, 20359 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8967-9528","authenticated-orcid":false,"given":"Hagen","family":"Frickmann","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 20359 Hamburg, Germany"},{"name":"Institute for Medical Microbiology, Virology and Hygiene, University Medicine Rostock, 18057 Rostock, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ulrike","family":"Loderst\u00e4dt","sequence":"additional","affiliation":[{"name":"Department of Hospital Hygiene & Infectious Diseases, University Medicine G\u00f6ttingen, 37075 G\u00f6ttingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"21807","DOI":"10.1038\/s41598-020-78691-7","article-title":"Misidentification of Plasmodium ovale as Plasmodium vivax malaria by a microscopic method: A meta-analysis of confirmed P. ovale cases","volume":"10","author":"Kotepui","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"101442","DOI":"10.1016\/j.tmaid.2019.06.013","article-title":"Evaluation of the multiplex real-time PCR assays RealStar malaria S & T PCR kit 1.0 and FTD malaria differentiation for the differentiation of Plasmodium species in clinical samples","volume":"31","author":"Frickmann","year":"2019","journal-title":"Travel Med. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1186\/s12936-019-2901-0","article-title":"A comparison of two PCR protocols for the differentiation of Plasmodium ovale species and implications for clinical management in travellers returning to Germany: A 10-year cross-sectional study","volume":"18","author":"Frickmann","year":"2019","journal-title":"Malar. J."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.1111\/j.1469-0691.2009.02933.x","article-title":"Comparison of three real-time PCR methods with blood smears and rapid diagnostic test in Plasmodium sp. infection","volume":"16","author":"Bourgeois","year":"2010","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1111\/j.1469-0691.2010.03218.x","article-title":"Multiplex real-time PCR for the diagnosis of malaria: Correlation with microscopy","volume":"17","author":"Dormond","year":"2011","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1016\/j.ijid.2020.07.009","article-title":"Systematic review and meta-analysis of diagnostic accuracy of loop-mediated isothermal amplification (LAMP) methods compared with microscopy, polymerase chain reaction and rapid diagnostic tests for malaria diagnosis","volume":"98","author":"Picot","year":"2020","journal-title":"Int. J. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1128\/JCM.01858-08","article-title":"Multiplexed real-time PCR assay for discrimination of Plasmodium species with improved sensitivity for mixed infections","volume":"47","author":"Shokoples","year":"2009","journal-title":"J. Clin. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1007\/s10096-016-2844-0","article-title":"Contribution of real-time PCR to Plasmodium species identification and to clinical decisions: A nationwide study in a non-endemic setting","volume":"36","author":"Grossman","year":"2017","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"12765","DOI":"10.1038\/s41598-020-69647-y","article-title":"Summary of discordant results between rapid diagnosis tests, microscopy, and polymerase chain reaction for detecting Plasmodium mixed infection: A systematic review and meta-analysis","volume":"10","author":"Kotepui","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1186\/s12936-021-03783-2","article-title":"Assessing field performance of ultrasensitive rapid diagnostic tests for malaria: A systematic review and meta-analysis","volume":"20","author":"Danwang","year":"2021","journal-title":"Malar. J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e01508-20","DOI":"10.1128\/JCM.01508-20","article-title":"Ultrasensitive Diagnostics for Low-Density Asymptomatic Plasmodium falciparum Infections in Low-Transmission Settings","volume":"59","author":"Zainabadi","year":"2021","journal-title":"J. Clin. Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e1387","DOI":"10.1093\/cid\/ciab221","article-title":"The role of ultra-sensitive molecular methods for detecting malaria\u2014The broader perspective","volume":"7","author":"Kamaliddin","year":"2021","journal-title":"Clin. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"104987","DOI":"10.1016\/j.meegid.2021.104987","article-title":"Asymptomatic malaria infection prevailing risks for human health and malaria elimination","volume":"93","author":"Prusty","year":"2021","journal-title":"Infect. Genet. Evol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Mbanefo, A., and Kumar, N. (2020). Evaluation of Malaria Diagnostic Methods as a Key for Successful Control and Elimination Programs. Trop. Med. Infect. Dis., 5.","DOI":"10.3390\/tropicalmed5020102"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e366","DOI":"10.1016\/S2666-5247(21)00055-0","article-title":"Global patterns of submicroscopic Plasmodium falciparum malaria infection: Insights from a systematic review and meta-analysis of population surveys","volume":"2","author":"Whittaker","year":"2021","journal-title":"Lancet Microbe"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1186\/s12936-021-03667-5","article-title":"What is the yield of malaria reactive case detection in the Greater Mekong Sub-region? A review of published data and meta-analysis","volume":"20","author":"Deen","year":"2021","journal-title":"Malar. J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1296","DOI":"10.1093\/infdis\/jiy319","article-title":"Use of Capillary Blood Samples Leads to Higher Parasitemia Estimates and Higher Diagnostic Sensitivity of Microscopic and Molecular Diagnostics of Malaria Than Venous Blood Samples","volume":"218","author":"Mischlinger","year":"2018","journal-title":"J. Infect. Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e005634","DOI":"10.1136\/bmjgh-2021-005634","article-title":"Accuracy of malaria diagnostic tests performed on non-invasively collected samples: A systematic review and meta-analysis","volume":"6","author":"Danwang","year":"2021","journal-title":"BMJ Glob. Health"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4845","DOI":"10.1038\/s41598-021-83977-5","article-title":"Comparative performance of PCR using DNA extracted from dried blood spots and whole blood samples for malaria diagnosis: A meta-analysis","volume":"11","author":"Mahittikorn","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1186\/1475-2875-12-106","article-title":"Rapid diagnostic tests for molecular surveillance of Plasmodium falciparum malaria -assessment of DNA extraction methods and field applicability","volume":"12","author":"Morris","year":"2013","journal-title":"Malar. J."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1186\/1475-2875-9-370","article-title":"Giemsa-stained thick blood films as a source of DNA for Plasmodium species-specific real-time PCR","volume":"9","author":"Cnops","year":"2010","journal-title":"Malar. J."},{"key":"ref_22","first-page":"634","article-title":"Detection of Plasmodium spp. in human feces","volume":"18","year":"2012","journal-title":"Emerg. Inf. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Loderst\u00e4dt, U., Hagen, R.M., Hahn, A., and Frickmann, H. (2021). New Developments in PCR-Based Diagnostics for Bacterial Pathogens Causing Gastrointestinal Infections-A Narrative Mini-Review on Challenges in the Tropics. Trop. Med. Infect. Dis., 6.","DOI":"10.3390\/tropicalmed6020096"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"616","DOI":"10.3109\/00365548.2013.777777","article-title":"Increased detection of invasive enteropathogenic bacteria in pre-incubated blood culture materials by real-time PCR in comparison with automated incubation in Sub-Saharan Africa","volume":"45","author":"Frickmann","year":"2013","journal-title":"Scand. J. Infect. Dis."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Frickmann, H., Dekker, D., Schwarz, N.G., Hahn, A., Boahen, K., Sarpong, N., Adu-Sarkodie, Y., Halbgewachs, E., Marks, F., and von Kalckreuth, V. (2015). 16S rRNA Gene Sequence-Based Identification of Bacteria in Automatically Incubated Blood Culture Materials from Tropical Sub-Saharan Africa. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0135923"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Nielsen, M.V., Sarpong, N., Krumkamp, R., Dekker, D., Loag, W., Amemasor, S., Agyekum, A., Marks, F., Huenger, F., and Krefis, A.C. (2012). Incidence and characteristics of bacteremia among children in rural Ghana. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0044063"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1796","DOI":"10.3201\/eid1611.100388","article-title":"Typhoid fever among children, Ghana","volume":"16","author":"Marks","year":"2010","journal-title":"Emerg. Infect. Dis."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Schwarz, N.G., Sarpong, N., H\u00fcnger, F., Marks, F., Acquah, S.E., Agyekum, A., Nkrumah, B., Loag, W., Hagen, R.M., and Evans, J.A. (2010). Systemic bacteraemia in children presenting with clinical pneumonia and the impact of non-typhoid salmonella (NTS). BMC Infect. Dis., 10.","DOI":"10.1186\/1471-2334-10-319"},{"key":"ref_29","first-page":"307","article-title":"Eine Vereinfachung und Vervollkommnung meiner Methylenblau-Eosin-F\u00e4rbemethode zur Erzielung der Romanowsky-Nocht\u2019schen Chromatinf\u00e4rbung","volume":"32","author":"Giemsa","year":"1904","journal-title":"Cent. F\u00fcr Bakteriol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/0035-9203(85)90329-3","article-title":"Rapid evaluation of malaria parasite density and standardization of thick smear examination for epidemiological investigations","volume":"79","author":"Trape","year":"1985","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2435","DOI":"10.1128\/JCM.43.5.2435-2440.2005","article-title":"Real-time PCR for detection and identification of Plasmodium spp.","volume":"43","author":"Mangold","year":"2005","journal-title":"J. Clin. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1515\/labmed-2019-0174","article-title":"Meta-analysis of the diagnostic performance characteristics of three commercial and one in-house nucleic acid amplification tests for malaria screening","volume":"44","author":"Altangerel","year":"2020","journal-title":"J. Lab. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1186\/1475-2875-11-307","article-title":"Real-time PCR assay for discrimination of Plasmodium ovale curtisi and Plasmodium ovale wallikeri in the Ivory Coast and in the Comoros Islands","volume":"11","author":"Bauffe","year":"2012","journal-title":"Malar. J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1624","DOI":"10.1128\/JCM.02316-06","article-title":"Genetic polymorphisms influence Plasmodium ovale PCR detection accuracy","volume":"45","author":"Calderaro","year":"2007","journal-title":"J. Clin. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Calderaro, A., Piccolo, G., Gorrini, C., Montecchini, S., Rossi, S., Medici, M.C., Chezzi, C., and Snounou, G. (2012). A new real-time PCR for the detection of Plasmodium ovale wallikeri. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0048033"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"h5527","DOI":"10.1136\/bmj.h5527","article-title":"STARD 2015: An updated list of essential items for reporting diagnostic accuracy studies","volume":"351","author":"Bossuyt","year":"2015","journal-title":"BMJ"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1186\/s12879-021-06290-6","article-title":"Comparison of capillary, venous and buffy coat blood samples in detecting Plasmodium species among malaria suspected patients attending at Hamusite health center. A cross-sectional study","volume":"21","author":"Abeje","year":"2021","journal-title":"BMC Infect. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1186\/1475-2875-10-380","article-title":"Plasmodium falciparum population dynamics during the early phase of anti-malarial drug treatment in Tanzanian children with acute uncomplicated malaria","volume":"10","author":"Carlsson","year":"2011","journal-title":"Malar. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"105377","DOI":"10.1016\/j.actatropica.2020.105377","article-title":"On detection thresholds-a review on diagnostic approaches in the infectious disease laboratory and the interpretation of their results","volume":"205","author":"Hahn","year":"2020","journal-title":"Acta Trop."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1186\/s12936-021-03618-0","article-title":"High frequency of the Duffy-negative genotype and absence of Plasmodium vivax infections in Ghana","volume":"20","author":"Brown","year":"2021","journal-title":"Malar. J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1093\/oxfordjournals.aje.a112510","article-title":"Estimating prevalence from the results of a screening test","volume":"107","author":"Rogan","year":"1978","journal-title":"Am. J. Epidemiol."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/10\/2\/406\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:17:40Z","timestamp":1760134660000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/10\/2\/406"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,10]]},"references-count":41,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["microorganisms10020406"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms10020406","relation":{},"ISSN":["2076-2607"],"issn-type":[{"type":"electronic","value":"2076-2607"}],"subject":[],"published":{"date-parts":[[2022,2,10]]}}}