{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T02:02:20Z","timestamp":1785290540575,"version":"3.55.0"},"reference-count":33,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T00:00:00Z","timestamp":1776038400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T00:00:00Z","timestamp":1776038400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transfusion"],"published-print":{"date-parts":[[2026,6]]},"abstract":"Abstract<\/jats:title>\n \n Background<\/jats:title>\n \n Anaplasma phagocytophilum<\/jats:italic>\n is a tick\u2010borne pathogen that causes anaplasmosis. Increased incidences of this disease in Canada and cases of transfusion\u2010transmitted anaplasmosis in the United States have been reported. Currently, there are no Health Canada licensed methodologies available for detection of\n A. phagocytophilum<\/jats:italic>\n nucleic acids (NAs) in blood donors. Here we describe a method for NA extraction and detection of\n A. phagocytophilum<\/jats:italic>\n in whole blood (WB) donor samples.\n <\/jats:p>\n <\/jats:sec>\n \n Study Design and Methods<\/jats:title>\n \n Plasmids carrying the\n A. phagocytophilum msp2<\/jats:italic>\n and\n gltA<\/jats:italic>\n genes and genomic\n A. phagocytophilum<\/jats:italic>\n DNA were used to spike WB (10\n 2<\/jats:sup>\n \u201310\n 10<\/jats:sup>\n plasmid copies, and 3.35\u2009\u00d7\u200910\n 6<\/jats:sup>\n genome copies\/1.1\u2009mL of WB, respectively) which was added to proprietary WB collection tubes (RWBCT). The tubes were frozen at \u221220\u00b0C for \u226548\u2009h followed by NA extraction and detection of the\n msp2<\/jats:italic>\n and\n gltA<\/jats:italic>\n genes using SYBR green\u2010based qPCR. Unspiked WB and human glyceraldehyde\u20103\u2010phosphate dehydrogenase (GAPDH) gene amplification were used as process controls. Specificity was assessed with plasmids containing gene fragments from\n Anaplasma marginale<\/jats:italic>\n and\n Anaplasma platys<\/jats:italic>\n . Assays were run in triplicates.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n A. phagocytophilum msp2<\/jats:italic>\n and\n gltA<\/jats:italic>\n were detected by qPCR in plasmid\u2010spiked WB at a concentration of 10\n 3<\/jats:sup>\n copies\/1.1\u2009mL of WB; however, the\n gltA<\/jats:italic>\n primer set yielded false positive results. Additionally, genomic\n A. phagocytophilum<\/jats:italic>\n DNA was successfully detected in spiked WB using\n msp2<\/jats:italic>\n specific primers. Cross reaction with\n A. platys msp2<\/jats:italic>\n and\n A. marginale gltA<\/jats:italic>\n was observed with the\n A. phagocytophilum<\/jats:italic>\n primer sets.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n A simple, scalable in\u2010house protocol for NA extraction from RWBCT has been optimized to detect\n A. phagocytophilum<\/jats:italic>\n in WB donor samples.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1111\/trf.70194","type":"journal-article","created":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T08:10:31Z","timestamp":1776067831000},"page":"1122-1129","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["An in\u2010house nucleic acid test for detection of\n Anaplasma phagocytophilum<\/i>\n in whole blood donor samples"],"prefix":"10.1111","volume":"66","author":[{"given":"Dilini","family":"Kumaran","sequence":"first","affiliation":[{"name":"Surveillance and Discovery Laboratory Canadian Blood Services Ottawa Ontario Canada"},{"name":"Medical Microbiology Canadian Blood Services Ottawa Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9129-5136","authenticated-orcid":false,"given":"Carmen","family":"Charlton","sequence":"additional","affiliation":[{"name":"Surveillance and Discovery Laboratory Canadian Blood Services Ottawa Ontario Canada"},{"name":"Laboratory Medicine and Pathology University of Alberta Edmonton Alberta Canada"},{"name":"Medical Microbiology Canadian Blood Services Edmonton Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chantale","family":"Pambrun","sequence":"additional","affiliation":[{"name":"Innovation & Portfolio Management Canadian Blood Services Ottawa Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2519-1109","authenticated-orcid":false,"given":"Steven J.","family":"Drews","sequence":"additional","affiliation":[{"name":"Surveillance and Discovery Laboratory Canadian Blood Services Ottawa Ontario Canada"},{"name":"Laboratory Medicine and Pathology University of Alberta Edmonton Alberta Canada"},{"name":"Medical Microbiology Canadian Blood Services Edmonton Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0705-4987","authenticated-orcid":false,"given":"Sandra","family":"Ramirez\u2010Arcos","sequence":"additional","affiliation":[{"name":"Surveillance and Discovery Laboratory Canadian Blood Services Ottawa Ontario Canada"},{"name":"Medical Microbiology Canadian Blood Services Ottawa Ontario Canada"},{"name":"Department of Biochemistry, Microbiology and Immunology University of Ottawa Ottawa Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,13]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1086\/518146"},{"key":"e_1_2_9_3_1","volume-title":"StatPearls [Internet]","author":"Guzman 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