{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T20:25:46Z","timestamp":1785356746776,"version":"3.55.0"},"reference-count":31,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2023,12,19]],"date-time":"2023-12-19T00:00:00Z","timestamp":1702944000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,5,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Objectives<\/jats:title>\n                  <jats:p>Recent work has demonstrated that automated fluorescence flow cytometry (FLC) is a potential alternative for the detection and quantification of Plasmodium parasites. The objective of this study was to apply this novel FLC method to detect and quantify Babesia parasites in venous blood and compare results to light microscopy and polymerase chain reaction methods.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>An automated hematology\/malaria analyzer (XN-31; Sysmex) was used to detect and quantify B microti\u2013infected red blood cells from residual venous blood samples (n = 250: Babesia positive, n = 170; Babesia negative, n = 80). As no instrument software currently exists for Babesia, qualitative and quantitative machine learning (ML) algorithms were developed to facilitate analysis.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Performance of the ML models was verified against the XN-31 software using P falciparum\u2013infected samples. When applied to Babesia-infected samples, the qualitative ML model demonstrated an area under the curve (AUC) of 0.956 (sensitivity, 95.9%; specificity, 83.3%) relative to polymerase chain reaction. For valid scattergrams, the qualitive model achieved an AUC of 1.0 (sensitivity and specificity, 100%), while the quantitative model demonstrated an AUC of 0.986 (sensitivity, 94.4%; specificity, 100%).<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>This investigation demonstrates that Babesia parasites can be detected and quantified directly from venous blood using FLC. Although promising, opportunities remain to improve the general applicability of the method.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/ajcp\/aqad168","type":"journal-article","created":{"date-parts":[[2023,12,19]],"date-time":"2023-12-19T19:51:09Z","timestamp":1703015469000},"page":"451-462","source":"Crossref","is-referenced-by-count":5,"title":["Detection and quantification of <i>Babesia<\/i> species intraerythrocytic parasites by flow cytometry"],"prefix":"10.1093","volume":"161","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1341-1733","authenticated-orcid":false,"given":"Patrick M","family":"Vanderboom","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anisha","family":"Misra","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9308-4810","authenticated-orcid":false,"given":"Kyle G","family":"Rodino","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Allison R","family":"Eberly","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason D","family":"Greenwood","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heather E","family":"Morris","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felicity C","family":"Norrie","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9300-4451","authenticated-orcid":false,"given":"Emily C","family":"Fernholz","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bobbi S","family":"Pritt","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2955-2066","authenticated-orcid":false,"given":"Andrew P","family":"Norgan","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic , Rochester, MN , US"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,12,19]]},"reference":[{"key":"2024050205534073400_CIT0001","first-page":"171","article-title":"Guidelines on the use of therapeutic apheresis in clinical practice\u2014evidence-based approach from the Writing Committee of the American Society for Apheresis: the eighth special issue","volume-title":"J Clin Apher.","author":"Padmanabhan","year":"2019"},{"key":"2024050205534073400_CIT0002","first-page":"185","article-title":"Clinical practice guidelines by the Infectious Diseases Society of America (IDSA): 2020 guideline on diagnosis and management of babesiosis","volume-title":"Clin Infect Dis","author":"Krause","year":"2021"},{"key":"2024050205534073400_CIT0003","first-page":"2149","article-title":"Human babesiosis in New York state: review of 139 hospitalized cases and analysis of prognostic factors archives of internal medicine","volume-title":"Arch Intern Med.","author":"White","year":"1998"},{"key":"2024050205534073400_CIT0004","first-page":"77","article-title":"Guidelines on the use of therapeutic apheresis in clinical practice\u2014evidence-based approach from the Writing Committee of the American Society for Apheresis: the ninth special issue","volume-title":"J Clin Apher.","author":"Connelly-Smith","year":"2023"},{"key":"2024050205534073400_CIT0005","first-page":"564","article-title":"Malaria: current status of control, diagnosis, treatment, and a proposed agenda for research and development","volume-title":"Lancet Infect Dis.","author":"Guerin","year":"2002"},{"key":"2024050205534073400_CIT0006","first-page":"233","article-title":"External quality assessment of malaria microscopy diagnosis in selected health facilities in Western Oromia, Ethiopia","volume-title":"Malar J.","author":"Sori","year":"2018"},{"key":"2024050205534073400_CIT0007","first-page":"218","article-title":"Applications of digital microscopy and densely connected convolutional neural networks for automated quantification of babesia-infected erythrocytes","volume-title":"Clin Chem.","author":"Durant","year":"2021"},{"issue":"2","key":"2024050205534073400_CIT0008","first-page":"311","article-title":"Imprecision of ratio-derived differential leukocyte counts","volume":"11","author":"R\u00fcmke","year":"1985","journal-title":"Blood Cells."},{"key":"2024050205534073400_CIT0009","first-page":"279","article-title":"Peripheral blood film review. The demise of the eyecount leukocyte differential","volume-title":"Clin Lab Med.","author":"Pierre","year":"2002"},{"key":"2024050205534073400_CIT0010","first-page":"2816","article-title":"Use of the quantitative buffy coat system for detection of parasitemia in patients with babesiosis","volume-title":"J Clin Microbiol.","author":"Mattia","year":"1993"},{"issue":"1","key":"2024050205534073400_CIT0011","first-page":"57","article-title":"Comparative study of peripheral blood smear and quantitative buffy coat in malaria diagnosis","volume":"43","author":"Salmani","year":"2011","journal-title":"J Commun Dis."},{"key":"2024050205534073400_CIT0012","first-page":"1838","article-title":"Comparison of microscopy, nested-PCR, and real-time-PCR assays using high-throughput screening of pooled samples for diagnosis of malaria in asymptomatic carriers from areas of endemicity in Myanmar","volume-title":"J Clin Microbiol.","author":"Wang","year":"2014"},{"key":"2024050205534073400_CIT0013","first-page":"1108","article-title":"Assessment of ultra-sensitive malaria diagnosis versus standard molecular diagnostics for malaria elimination: an in-depth molecular community cross-sectional study","volume-title":"Lancet Infect Dis.","author":"Hofmann","year":"2018"},{"key":"2024050205534073400_CIT0014","first-page":"446","article-title":"Malaria diagnosis for malaria elimination","volume-title":"Curr Opin Infect Dis.","author":"Zimmerman","year":"2015"},{"key":"2024050205534073400_CIT0015","volume-title":"Guidelines for the Treatment of Malaria","author":"World Health Organization","year":"2010","edition":"2"},{"key":"2024050205534073400_CIT0016","first-page":"100073","article-title":"Artificial intelligence for malaria: bridging the gap for real world usage","volume-title":"Lancet Reg Health Southeast Asia.","author":"Fan","year":"2022"},{"key":"2024050205534073400_CIT0017","first-page":"643","article-title":"Cytometry in malaria: moving beyond Giemsa","volume-title":"Cytometry A.","author":"Shapiro","year":"2007"},{"key":"2024050205534073400_CIT0018","first-page":"1067","article-title":"An adaptable two-color flow cytometric assay to quantitate the invasion of erythrocytes by Plasmodium falciparum parasites","volume-title":"Cytometry A","author":"Theron","year":"2010"},{"key":"2024050205534073400_CIT0019","first-page":"329","article-title":"Detection of antibodies to variant antigens on Plasmodium falciparum-infected erythrocytes by flow cytometry","volume-title":"Cytometry","author":"Staalsoe","year":"1999"},{"key":"2024050205534073400_CIT0020","first-page":"3116","article-title":"Impairment of Plasmodium falciparum growth in thalassemic red blood cells: further evidence by using biotin labeling and flow cytometry","volume-title":"Blood.","author":"Pattanapanyasat","year":"1999"},{"key":"2024050205534073400_CIT0021","first-page":"34","article-title":"Plasmodium vivax: isotopic, PicoGreen, and microscopic assays for measuring chloroquine sensitivity in fresh and cryopreserved isolates","volume-title":"Exp Parasitol.","author":"Kosaisavee","year":"2006"},{"key":"2024050205534073400_CIT0022","first-page":"215","article-title":"Analysis of Plasmodium falciparum growth in culture using acridine orange and flow cytometry","volume-title":"J Histochem Cytochem.","author":"Hare","year":"1986"},{"key":"2024050205534073400_CIT0023","first-page":"546","article-title":"Monitoring Plasmodium falciparum growth and development by UV flow cytometry using an optimized Hoechst-thiazole orange staining strategy","volume-title":"Cytometry A","author":"Grimberg","year":"2008"},{"key":"2024050205534073400_CIT0024","first-page":"59","article-title":"An automated haematology analyzer XN-30 distinguishes developmental stages of falciparum malaria parasite cultured in vitro","volume-title":"Malar J.","author":"Tougan","year":"2018"},{"key":"2024050205534073400_CIT0025","first-page":"5268","article-title":"Performance of the hematology analyzer XN-31 prototype in the detection of Plasmodium infections in an endemic region of Colombia","volume-title":"Sci Rep.","author":"Zuluaga-Id\u00e1rraga","year":"2021"},{"key":"2024050205534073400_CIT0026","first-page":"15","article-title":"Evaluation of automated malaria diagnosis using the Sysmex XN-30 analyser in a clinical setting","volume-title":"Malar J.","author":"Pillay","year":"2019"},{"key":"2024050205534073400_CIT0027","first-page":"231","article-title":"A method for reducing the sloughing of thick blood films for malaria diagnosis","volume-title":"Malar J.","author":"Norgan","year":"2013"},{"key":"2024050205534073400_CIT0028","article-title":"flowWorkspace: infrastructure for representing and interacting with gated and ungated cytometry data sets","author":"Finak","year":"2022"},{"key":"2024050205534073400_CIT0029","first-page":"Article25","article-title":"Super learner","volume-title":"Stat Appl Genet Mol Biol.","author":"van der Laan","year":"2007"},{"key":"2024050205534073400_CIT0030","first-page":"697669","article-title":"Babesia microti: pathogen genomics, genetic variability, immunodominant antigens, and pathogenesis","volume-title":"Front Microbiol.","author":"Puri","year":"2021"},{"key":"2024050205534073400_CIT0031","first-page":"498","article-title":"Genome sequence of the human malaria parasite Plasmodium falciparum.","volume-title":"Nature.","author":"Gardner","year":"2002"}],"container-title":["American Journal of Clinical Pathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/ajcp\/article-pdf\/161\/5\/451\/57348390\/aqad168.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/ajcp\/article-pdf\/161\/5\/451\/57348390\/aqad168.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,2]],"date-time":"2024-05-02T05:56:05Z","timestamp":1714629365000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/ajcp\/article\/161\/5\/451\/7479640"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,19]]},"references-count":31,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2023,12,19]]},"published-print":{"date-parts":[[2024,5,2]]}},"URL":"https:\/\/doi.org\/10.1093\/ajcp\/aqad168","relation":{},"ISSN":["0002-9173","1943-7722"],"issn-type":[{"value":"0002-9173","type":"print"},{"value":"1943-7722","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2024,5,1]]},"published":{"date-parts":[[2023,12,19]]}}}