{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T04:04:04Z","timestamp":1782446644842,"version":"3.54.5"},"reference-count":28,"publisher":"Frontiers Media SA","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\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Sports Act. Living"],"abstract":"<jats:p>This article presents the results of a study aimed to give new suggestions and strategies for improving the implementation of the flow cytofluorimetry-based method for the detection of homologous blood transfusions in doping control. The method is based on the recognition of the phenotypic mismatch between minority blood group antigens possessed by the donor and the recipient. Two strategies have been followed to reduce the risk of false-negative results: (i) the monitoring of a broader range of erythrocytes surface antigens; and (ii) the application of different surface erythrocyte staining protocols, tailored on the different antigens and the type of antigenic mismatch that had to be detected (whether it is the donor or the recipient who expresses or not the antigen to be detected). Special attention has also been focused on the time factor, to avoid prolonged sample storage, since hemolysis may have a significant impact on the reliability and quality of the results. Our experimental evidence suggests that the risk of false-negative results can be minimized by (i) the expansion of the antigen panel, with the inclusion of four additional targets; (ii) a more accurate selection of the gating area of the red blood cells; (iii) the choice of a better fluorochrome (alexa fluor 488) to be conjugated to the secondary antibody; and (iv) the implementation of different staining protocols depending on the nature of the double population to be detected (donor expressing vs. recipient non-expressing and vice versa). The combination of the above approaches allowed a significant reduction of false-negative results, assessed on samples simulating a homologous blood transfusion between two compatible subjects.<\/jats:p>","DOI":"10.3389\/fspor.2022.808449","type":"journal-article","created":{"date-parts":[[2022,2,10]],"date-time":"2022-02-10T06:35:35Z","timestamp":1644474935000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":15,"title":["Detection of Homologous Blood Transfusion in Sport Doping by Flow Cytofluorimetry: State of the Art and New Approaches to Reduce the Risk of False-Negative Results"],"prefix":"10.3389","volume":"4","author":[{"given":"Francesco","family":"Donati","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xavier","family":"de la Torre","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarajane","family":"Pagliarosi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Pirri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuliana","family":"Prevete","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Botr\u00e8","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,2,10]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"657","DOI":"10.1002\/ajh.21196","article-title":"Blood doping in athletes\u2013detection of allogeneic blood transfusions by flow cytofluorometry","volume":"83","author":"Arndt","year":"2008","journal-title":"Am. J. Hematol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"2379","DOI":"10.1002\/rcm.523","article-title":"RSR13, a potential athletic performance enhancement agent: detection in urine by gas chromatography\/mass spectrometry","volume":"15","author":"Breidbach","year":"2001","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"B3","article-title":"Homologous Blood transfusion (HBT) test by cytometry for Chinese and non-Chinese population","volume-title":"Recent Advances in Doping Analysis (17) - Proceedings of the 27th Cologne Workshop","author":"Chunji","year":"2009"},{"key":"B4","doi-asserted-by":"publisher","first-page":"250","DOI":"10.1111\/j.1423-0410.2007.00887.x","article-title":"International Society of Blood Transfusion Committee on terminology for red cell surface antigens: Cape Town report","volume":"92","author":"Daniels","year":"2007","journal-title":"Vox Sang"},{"key":"B5","doi-asserted-by":"publisher","first-page":"252","DOI":"10.1016\/j.talanta.2009.11.066","article-title":"A fast screening method for the detection of the abuse of hemoglobin-based oxygen carriers (HBOCs) in doping control","volume":"81","author":"Donati","year":"2010","journal-title":"Talanta"},{"key":"B6","doi-asserted-by":"publisher","first-page":"2915","DOI":"10.1002\/elps.201100178","article-title":"A simple and rapid pre-confirmation method to distinguish endogenous human haemoglobin from synthetic haemoglobin-based oxygen carriers in doping control","volume":"32","author":"Donati","year":"2011","journal-title":"Electrophoresis"},{"key":"B7","doi-asserted-by":"publisher","first-page":"2276","DOI":"10.1096\/fasebj.2021.35.S1.02276","article-title":"Improving the detection of homologous blood transfusion in sport doping: a full DNA-based genotyping strategy on dried blood spots","volume":"35","author":"Donati","year":"2021","journal-title":"FASEB J"},{"key":"B8","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1016\/j.talanta.2013.02.042","article-title":"Investigation on the application of DNA forensic human identification techniques to detect homologous blood transfusions in doping control","volume":"110","author":"Donati","year":"2013","journal-title":"Talanta"},{"key":"B9","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/j.forsciint.2008.04.007","article-title":"Scientific and forensic standards for homologous blood transfusion anti-doping analyses","volume":"179","author":"Giraud","year":"2008","journal-title":"Forensic Sci. Int."},{"key":"B10","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1007\/978-3-540-79088-4_13","article-title":"Blood transfusion in sports","volume":"2010","author":"Giraud","year":"2010","journal-title":"Handb. Exp. Pharmacol"},{"key":"B11","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1093\/chromsci\/43.1.39","article-title":"Methodologies for detection of hemoglobin-based oxygen carriers","volume":"43","author":"Goebel","year":"2005","journal-title":"J Chromatogr Sci."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1160","DOI":"10.1002\/dta.1736","article-title":"Possible cause of lack of positive samples on homologous blood transfusion","volume":"6","author":"Krotov","year":"2014","journal-title":"Drug Test Anal"},{"key":"B13","doi-asserted-by":"publisher","first-page":"954","DOI":"10.1016\/j.jpba.2009.06.038","article-title":"A high-throughput test to detect C.E.R.A. doping in blood","volume":"50","author":"Lamon","year":"2009","journal-title":"J. Pharm. Biomed. Anal."},{"key":"B14","doi-asserted-by":"publisher","first-page":"635","DOI":"10.1038\/35015164","article-title":"Recombinant erythropoietin in urine","volume":"405","author":"Lasne","year":"2000","journal-title":"Nature"},{"key":"B15","doi-asserted-by":"publisher","first-page":"888","DOI":"10.3324\/haematol.2009.006809","article-title":"Detection of continuous erythropoietin receptor activator in blood and urine in anti-doping control","volume":"94","author":"Lasne","year":"2009","journal-title":"Haematologica"},{"key":"B16","doi-asserted-by":"publisher","first-page":"728273","DOI":"10.3389\/fmolb.2021.728273","article-title":"Integrating whole blood transcriptomic collection procedures into the current anti-doping testing system, including long-term storage and re-testing of anti-doping samples","volume":"8","author":"Lima","year":"2021","journal-title":"Front. Mol. Biosci"},{"key":"B17","doi-asserted-by":"publisher","first-page":"9693","DOI":"10.1007\/s00216-013-7122-8","article-title":"A DNA-based method for detecting homologous blood doping","volume":"405","author":"Manokhina","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"B18","volume-title":"Human Biology and Health","author":"Maton","year":"1993"},{"key":"B19","doi-asserted-by":"publisher","first-page":"3024","DOI":"10.1016\/j.jchromb.2010.08.046","article-title":"Microwave irradiation for a fast gas chromatography-mass spectrometric analysis of polysaccharide-based plasma volume expanders in human urine","volume":"878","author":"Mazzarino","year":"2010","journal-title":"J. Chromatogr. B."},{"key":"B20","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1111\/j.1537-2995.2009.02352.x","article-title":"Urinary di-(2-ethylhexyl)phthalate metabolites in athletes as screening measure for illicit blood doping: a comparison study with patients receiving blood transfusion","volume":"50","author":"Monfort","year":"2010","journal-title":"Transfusion"},{"key":"B21","doi-asserted-by":"publisher","first-page":"1284","DOI":"10.3324\/%25x","article-title":"Proof of homologous blood transfusion through quantification of blood group antigens","volume":"88","author":"Nelson","year":"2003","journal-title":"Haematologica"},{"key":"B22","doi-asserted-by":"publisher","first-page":"827","DOI":"10.1136\/bjsports-2014-093512","article-title":"Monitoring of biological markers indicative of doping: the athlete biological passport","volume":"48","author":"Saugy","year":"2014","journal-title":"Br. J. Sports Med."},{"key":"B23","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1016\/j.aca.2006.10.017","article-title":"Electrophoretic, size-exclusion high-performance liquid chromatography and liquid chromatography-electrospray ionization ion trap mass spectrometric detection of hemoglobin-based oxygen carriers","volume":"583","author":"Simitsek","year":"2007","journal-title":"Anal. Chim. Acta."},{"key":"B24","doi-asserted-by":"publisher","first-page":"204","DOI":"10.1016\/j.forsciint.2016.04.021","article-title":"Application of DNA-based forensic analysis for the detection of homologous transfusion of whole blood and of red blood cell concentrates in doping control","volume":"265","author":"Stampella","year":"2016","journal-title":"Forensic Sci. Int."},{"key":"B25","doi-asserted-by":"publisher","first-page":"1241","DOI":"10.1002\/elps.200900513","article-title":"Analysis of hemoglobin-based oxygen carriers by CE-UV\/Vis and CE-ESI-TOF\/MS","volume":"31","author":"Staub","year":"2010","journal-title":"Electrophoresis"},{"key":"B26","unstructured":"Anti-Doping Rule Violations - Olympic Games Tokyo2020"},{"key":"B27","doi-asserted-by":"publisher","first-page":"345","DOI":"10.1016\/S0378-4347(00)00251-6","article-title":"Detection of the plasma volume expander hydroxyethyl starch in human urine","volume":"744","author":"Thevis","year":"2000","journal-title":"J. Chromatogr. B."},{"key":"B28","doi-asserted-by":"publisher","first-page":"633","DOI":"10.1055\/s-2007-965076","article-title":"Detection of homologous blood transfusion","volume":"28","author":"Voss","year":"2007","journal-title":"Int J Sports Med."}],"container-title":["Frontiers in Sports and Active Living"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fspor.2022.808449\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,10]],"date-time":"2022-02-10T06:35:40Z","timestamp":1644474940000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fspor.2022.808449\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,10]]},"references-count":28,"alternative-id":["10.3389\/fspor.2022.808449"],"URL":"https:\/\/doi.org\/10.3389\/fspor.2022.808449","relation":{},"ISSN":["2624-9367"],"issn-type":[{"value":"2624-9367","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,10]]},"article-number":"808449"}}