{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,13]],"date-time":"2026-06-13T14:15:50Z","timestamp":1781360150250,"version":"3.54.1"},"reference-count":42,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2013,11,17]],"date-time":"2013-11-17T00:00:00Z","timestamp":1384646400000},"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":[[2014,5]]},"abstract":"Background<\/jats:title>Quantitation of fetomaternal hemorrhage (FMH<\/jats:styled-content>) is performed to determine the dose of prophylactic anti\u2010D<\/jats:styled-content> (RhIG<\/jats:styled-content>) required to prevent D<\/jats:styled-content> immunization of D<\/jats:styled-content>\u2013 women. Flow cytometry (FC<\/jats:styled-content>) is the most accurate method. However, maternal white blood cells (WBCs<\/jats:styled-content>) can give high background by binding anti\u2010D<\/jats:styled-content> nonspecifically, compromising accuracy.<\/jats:p><\/jats:sec>Study Design and Methods<\/jats:title>Maternal blood samples (69) were sent for FC<\/jats:styled-content> quantitation of FMH<\/jats:styled-content> after positive K<\/jats:styled-content>leihauer\u2010B<\/jats:styled-content>etke test (KBT<\/jats:styled-content>) analysis and RhIG<\/jats:styled-content> administration. Reagents used were BRAD<\/jats:styled-content>\u20103\u2013fluorescein isothiocyanate (FITC<\/jats:styled-content>; anti\u2010D<\/jats:styled-content>), AEVZ<\/jats:styled-content>5.3\u2010FITC<\/jats:styled-content> (anti\u2013varicella zoster [anti\u2010VZ<\/jats:styled-content>], negative control), anti\u2010fetal hemoglobin (HbF<\/jats:styled-content>)\u2010FITC<\/jats:styled-content>, blended two\u2010color reagents, BRAD<\/jats:styled-content>\u20103\u2010FITC<\/jats:styled-content>\/anti\u2010CD<\/jats:styled-content>45\u2010phycoerythrin (PE<\/jats:styled-content>; anti\u2010D<\/jats:styled-content>\/L<\/jats:styled-content>), and BRAD<\/jats:styled-content>\u20103\u2010FITC<\/jats:styled-content>\/anti\u2010CD<\/jats:styled-content>66b\u2010PE<\/jats:styled-content> (anti\u2010D<\/jats:styled-content>\/G<\/jats:styled-content>). PE<\/jats:styled-content>\u2010positive WBCs<\/jats:styled-content> were eliminated from analysis by gating. Full blood counts were performed on maternal samples and female donors.<\/jats:p><\/jats:sec>Results<\/jats:title>Elevated numbers of neutrophils were present in 80% of patients. Red blood cell (RBC<\/jats:styled-content>) indices varied widely in maternal blood. D<\/jats:styled-content>+ FMH<\/jats:styled-content> values obtained with anti\u2010D<\/jats:styled-content>\/L<\/jats:styled-content>, anti\u2010D<\/jats:styled-content>\/G<\/jats:styled-content>, and anti\u2010HbF<\/jats:styled-content>\u2010FITC<\/jats:styled-content> were very similar (r\u2009=\u20090.99, p\u2009<\u20090.001). Correlation between KBT<\/jats:styled-content> and anti\u2010HbF<\/jats:styled-content>\u2010FITC FMH<\/jats:styled-content> results was low (r\u2009=\u20090.716). Inaccurate FMH<\/jats:styled-content> quantitation using the current method (anti\u2010D<\/jats:styled-content> minus anti\u2010VZ<\/jats:styled-content>) occurred with 71% samples having less than 15\u2009mL of D<\/jats:styled-content>+ FMH<\/jats:styled-content> (RBCs<\/jats:styled-content>) and insufficient RhIG<\/jats:styled-content> calculated for 9%. Using two\u2010color reagents and anti\u2010HbF<\/jats:styled-content>\u2010FITC<\/jats:styled-content>, approximately 30% patients had elevated F<\/jats:styled-content> cells, 26% had no fetal cells, 6% had D<\/jats:styled-content>\u2013 FMH<\/jats:styled-content>, 26% had 4 to 15\u2009mL of D<\/jats:styled-content>+ FMH<\/jats:styled-content>, and 12% patients had more than 15\u2009mL of D<\/jats:styled-content>+ FMH<\/jats:styled-content> (RBCs) requiring more than 300\u2009\u03bcg of RhIG<\/jats:styled-content>.<\/jats:p><\/jats:sec>Conclusion<\/jats:title>Without accurate quantitation of D<\/jats:styled-content>+ FMH<\/jats:styled-content> by FC<\/jats:styled-content>, some women would receive inappropriate or inadequate anti\u2010D<\/jats:styled-content> prophylaxis. The latter may be at risk of immunization leading to hemolytic disease of the newborn.<\/jats:p><\/jats:sec>","DOI":"10.1111\/trf.12484","type":"journal-article","created":{"date-parts":[[2013,11,18]],"date-time":"2013-11-18T04:31:20Z","timestamp":1384749080000},"page":"1305-1316","source":"Crossref","is-referenced-by-count":9,"title":["Accurate quantitation of D<\/scp>+ fetomaternal hemorrhage by flow cytometry using a novel reagent to eliminate granulocytes from analysis"],"prefix":"10.1111","volume":"54","author":[{"given":"Belinda","family":"Kumpel","sequence":"first","affiliation":[{"name":"Bristol Institute for Transfusion Sciences NHS Blood and Transplant Bristol UK"},{"name":"International Blood Group Reference Laboratory NHS Blood and Transplant Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew","family":"Hazell","sequence":"additional","affiliation":[{"name":"Bristol Institute for Transfusion Sciences NHS Blood and Transplant Bristol UK"},{"name":"International Blood Group Reference Laboratory NHS Blood and Transplant Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan","family":"Guest","sequence":"additional","affiliation":[{"name":"International Blood Group Reference Laboratory NHS Blood and Transplant Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan","family":"Dixey","sequence":"additional","affiliation":[{"name":"International Blood Group Reference Laboratory NHS Blood and Transplant Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosey","family":"Mushens","sequence":"additional","affiliation":[{"name":"International Blood Group Reference Laboratory NHS Blood and Transplant Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Debbie","family":"Bishop","sequence":"additional","affiliation":[{"name":"Red Cell Immunohaematology Laboratory NHS Blood and Transplant Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tim","family":"Wreford\u2010Bush","sequence":"additional","affiliation":[{"name":"Blood Transfusion Laboratory Southmead Hospital Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edmond","family":"Lee","sequence":"additional","affiliation":[{"name":"Red Cell Immunohaematology Laboratory NHS Blood and Transplant London UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2013,11,17]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1537-2995.2008.01832.x"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.1990.30490273444.x"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0002-9378(87)90228-6"},{"key":"e_1_2_8_5_1","unstructured":"ParkerJ WrayJ GoochA et\u2009al.Guidelines for the use of prophylactic anti\u2010D immunoglobulin.2006. [cited 2012 Feb 14]. Available from:http:\/\/www.bcshguidelines.com"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0020-7292(99)90000-0"},{"key":"e_1_2_8_7_1","first-page":"43","volume-title":"Standards for blood banks and transfusion services. Rh immune globulin","author":"American Association of Blood Banks","year":"2012"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(59)92264-0"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-3148.2012.01143.x"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/tme.12030"},{"key":"e_1_2_8_11_1","doi-asserted-by":"crossref","first-page":"92","DOI":"10.21307\/immunohematology-2019-208","article-title":"Laboratory methods for Rh immunoprophylaxis: a review","volume":"26","author":"Sandler SG","year":"2010","journal-title":"Immunohematology"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.1982.22683068604.x"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1309\/3U97V7327JBDPAYN"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1093\/ajcp\/91.3.288"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.1998.38898375514.x"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.1996.36596282587.x"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.5858\/133.3.465"},{"key":"e_1_2_8_18_1","unstructured":"AustinE BatesS deSilvaM et\u2009al.Guidelines for the estimation of fetomaternal haemorrhage.2009. [cited 2009 Oct 10]. Available from:http:\/\/www.bcshguidelines.com"},{"issue":"1","key":"e_1_2_8_19_1","first-page":"33","article-title":"Use of a negative control antibody in the quantitation of feto\u2010maternal haemorrhage by flow cytometry","volume":"9","author":"Lloyd\u2010Evans P","year":"1999","journal-title":"Transfus Med"},{"issue":"1","key":"e_1_2_8_20_1","first-page":"26","article-title":"The importance of appropriate analysis of flow cytometry data in the estimation of fetomaternal haemorrhage","volume":"13","author":"Simpson E","year":"2003","journal-title":"Transfus Med"},{"key":"e_1_2_8_21_1","unstructured":"BaxterH LakumT leMaitreC.Assessment of fetomaternal haemorrhage: alternative flow cytometric techniques.The Biomedical Scientist2010;February:96\u201097."},{"key":"e_1_2_8_22_1","first-page":"2","article-title":"Estimation of fetomaternal haemorrhage using a flow cytometric technique","author":"Ward RY","year":"2011","journal-title":"ICCS E\u2010Newsletter"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.2000.40111376.x"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1537-2995.2011.03258.x"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(86)90837-8"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1423-0410.1990.tb02111.x"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj0850466"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj0880435"},{"key":"e_1_2_8_29_1","first-page":"227","article-title":"Factors affecting the antigen\u2010antibody reaction","volume":"5","author":"Reverberi R","year":"2007","journal-title":"Blood Transfus"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1080\/003655101753267937"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajog.2003.08.027"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.2001.41111445.x"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1423-0410.2011.01533.x"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1751-2824.2012.01568.x"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.2009.08054.x"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1136\/bmj.3.5817.31"},{"key":"e_1_2_8_37_1","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1182\/blood.V23.5.621.621","article-title":"Mechanisms of isoimmunization. I. The transplacental passage of fetal erythrocytes in homospecific pregnancies","volume":"23","author":"Cohen F","year":"1964","journal-title":"Blood"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(73)91675-9"},{"issue":"1","key":"e_1_2_8_39_1","first-page":"247","article-title":"Persistence of fetal haemoglobin as a cause of false positive Kleihauer\u2010Betke (acid elution) tests and excessive prophylactic anti\u2010D administration","volume":"105","author":"Hazell MJ","year":"2013","journal-title":"Vox Sang"},{"key":"e_1_2_8_40_1","article-title":"False positive Kleihauer\u2010Betke (acid elution) test caused by elevated maternal fetal haemoglobin F cells","author":"Donnelly JC","journal-title":"Europ J Obstet Gynecol Reprod Biol"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.1966.tb05636.x"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/0002-9378(86)90725-8"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-0528.2009.02244.x"}],"container-title":["Transfusion"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Ftrf.12484","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/trf.12484","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,29]],"date-time":"2023-09-29T15:42:46Z","timestamp":1696002166000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/trf.12484"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,11,17]]},"references-count":42,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2014,5]]}},"alternative-id":["10.1111\/trf.12484"],"URL":"https:\/\/doi.org\/10.1111\/trf.12484","archive":["Portico"],"relation":{},"ISSN":["0041-1132","1537-2995"],"issn-type":[{"value":"0041-1132","type":"print"},{"value":"1537-2995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,11,17]]}}}