{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,4]],"date-time":"2026-04-04T18:50:25Z","timestamp":1775328625353,"version":"3.50.1"},"reference-count":12,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2013,1,30]],"date-time":"2013-01-30T00:00:00Z","timestamp":1359504000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Transfusion"],"published-print":{"date-parts":[[2013,10]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\"><jats:italic>Babesia microti<\/jats:italic><\/jats:styled-content>, the most frequently implicated pathogen in transfusion\u2010transmitted babesiosis, is widely endemic in the <jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>ortheast and upper <jats:styled-content style=\"fixed-case\">M<\/jats:styled-content>idwestern <jats:styled-content style=\"fixed-case\">U<\/jats:styled-content>nited <jats:styled-content style=\"fixed-case\">S<\/jats:styled-content>tates. High seroprevalence in endemic areas limits antibody\u2010based donor screening. A high\u2010performance molecular test is needed to identify donors in the preseroconversion window phase as well as to discriminate past serologic exposure with parasite clearance from continued parasitemia.<\/jats:p><\/jats:sec><jats:sec><jats:title>Study Design and Methods<\/jats:title><jats:p>Frozen <jats:styled-content style=\"fixed-case\"><jats:italic>Babesia<\/jats:italic><\/jats:styled-content>\u2010spiked whole blood was microcentrifuged, and the supernatant transferred and microcentrifuged again to concentrate the parasite. The <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> was extracted and amplified using real\u2010time polymerase chain reaction (<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content>) using <jats:styled-content style=\"fixed-case\"><jats:italic>Babesia<\/jats:italic><\/jats:styled-content>\u2010specific primers. The assay was employed in three series of experiments: 1) a validation and optimization spiking experiment, 2) a blinded serial dilution probit analysis to determine the limit of detection, and 3) evaluation of two blinded panels of clinical samples from possible babesiosis cases.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>At a decreasing inoculum of 445, 44.5, and 4.45 copies\/<jats:styled-content style=\"fixed-case\">mL<\/jats:styled-content>, the assay had positive rates of 100, 97.5, and 81%, respectively. The blinded probit analysis demonstrated a detection rate of 95 and 50% at 12.92 and 1.52 parasites\/2\u2009<jats:styled-content style=\"fixed-case\">mL<\/jats:styled-content> of whole blood, respectively. Evaluation of clinical samples showed 13 of 21 samples to be positive, with a range of 85 to 4.8 million parasites\/<jats:styled-content style=\"fixed-case\">mL<\/jats:styled-content>. There were no positives detected among 48 healthy donors<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>We have developed a highly sensitive and specific, quantitative real\u2010time <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content>\u2010based assay for detection of <jats:styled-content style=\"fixed-case\"><jats:italic>B.\u2009microti<\/jats:italic><\/jats:styled-content> that could have a useful role in blood screening. It can also be employed broadly to understand <jats:styled-content style=\"fixed-case\"><jats:italic>Babesia<\/jats:italic><\/jats:styled-content> epidemiology, disease pathogenesis, and host immunology.<\/jats:p><\/jats:sec>","DOI":"10.1111\/trf.12098","type":"journal-article","created":{"date-parts":[[2013,1,30]],"date-time":"2013-01-30T11:53:23Z","timestamp":1359546803000},"page":"2299-2306","source":"Crossref","is-referenced-by-count":54,"title":["Development of a real\u2010time polymerase chain reaction assay for sensitive detection and quantitation of <scp><i>Babesia microti<\/i><\/scp> infection"],"prefix":"10.1111","volume":"53","author":[{"given":"Evan M.","family":"Bloch","sequence":"first","affiliation":[{"name":"Blood Systems Research Institute  San Francisco California"},{"name":"Yale School of Public Health and Yale School of Medicine  New Haven Connecticut"},{"name":"Tufts University Cummings School of Veterinary Medicine  North Grafton Massachusetts"},{"name":"Nantucket Cottage Hospital  Nantucket Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tzong\u2010Hae","family":"Lee","sequence":"additional","affiliation":[{"name":"Blood Systems Research Institute  San Francisco California"},{"name":"Yale School of Public Health and Yale School of Medicine  New Haven Connecticut"},{"name":"Tufts University Cummings School of Veterinary Medicine  North Grafton Massachusetts"},{"name":"Nantucket Cottage Hospital  Nantucket Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter J.","family":"Krause","sequence":"additional","affiliation":[{"name":"Blood Systems Research Institute  San Francisco California"},{"name":"Yale School of Public Health and Yale School of Medicine  New Haven Connecticut"},{"name":"Tufts University Cummings School of Veterinary Medicine  North Grafton Massachusetts"},{"name":"Nantucket Cottage Hospital  Nantucket Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"III","given":"Sam R.","family":"Telford","sequence":"additional","affiliation":[{"name":"Blood Systems Research Institute  San Francisco California"},{"name":"Yale School of Public Health and Yale School of Medicine  New Haven Connecticut"},{"name":"Tufts University Cummings School of Veterinary Medicine  North Grafton Massachusetts"},{"name":"Nantucket Cottage Hospital  Nantucket Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lani","family":"Montalvo","sequence":"additional","affiliation":[{"name":"Blood Systems Research Institute  San Francisco California"},{"name":"Yale School of Public Health and Yale School of Medicine  New Haven Connecticut"},{"name":"Tufts University Cummings School of Veterinary Medicine  North Grafton Massachusetts"},{"name":"Nantucket Cottage Hospital  Nantucket Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Chafets","sequence":"additional","affiliation":[{"name":"Blood Systems Research Institute  San Francisco California"},{"name":"Yale School of Public Health and Yale School of Medicine  New Haven Connecticut"},{"name":"Tufts University Cummings School of Veterinary Medicine  North Grafton Massachusetts"},{"name":"Nantucket Cottage Hospital  Nantucket Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sahar","family":"Usmani\u2010Brown","sequence":"additional","affiliation":[{"name":"Blood Systems Research Institute  San Francisco California"},{"name":"Yale School of Public Health and Yale School of Medicine  New Haven Connecticut"},{"name":"Tufts University Cummings School of Veterinary Medicine  North Grafton Massachusetts"},{"name":"Nantucket Cottage Hospital  Nantucket Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Timothy 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