{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T16:09:55Z","timestamp":1790525395594,"version":"4.1.0"},"reference-count":50,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2021,3,24]],"date-time":"2021-03-24T00:00:00Z","timestamp":1616544000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vox Sanguinis"],"published-print":{"date-parts":[[2021,10]]},"abstract":"<jats:sec><jats:title>Background and objectives<\/jats:title><jats:p>The aim of the study was to optimize routine non\u2010invasive\u00a0prenatal detection of fetal <jats:italic>RHD<\/jats:italic> gene from plasma of RhD\u2010negative pregnant women (the median of gestational age was 25\u00a0weeks, range 10\u201338) to detect RhD materno\u2010fetal incompatibility and to avoid the redundant immunoprophylaxis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and methods<\/jats:title><jats:p>Initially only one exon of <jats:italic>RHD<\/jats:italic> gene (exon 10) was investigated in 281 plasma samples (144 verified after delivery), in the second phase three <jats:italic>RHD<\/jats:italic> exons (5, 7, 10) were analyzed in 246 samples of plasma and maternal genomic DNA (204 verified) by real\u2010time PCR method. Detection of Y\u2010chromosomal sequence <jats:italic>DYS\u201014<\/jats:italic> and five X\u2010chromosomal insertion\/deletion polymorphisms was used to confirm the fetal cfDNA detectability in plasma. Specific polymorphisms in <jats:italic>RHD<\/jats:italic> gene were detected by sequence\u2010specific primer PCR in nine samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>When only the <jats:italic>RHD<\/jats:italic> exon 10 was tested, 2\u00b78% of verified samples were false positive and 3\u00b75% false negative. With three <jats:italic>RHD<\/jats:italic> exons (5, 7, 10) and maternal genomic DNA testing, only one case was false negative (0\u00b75%). Nine samples were inconclusive due to <jats:italic>RHD<\/jats:italic>\u2010positive results in maternal genomic DNA. These samples were analyzed for specific mutations in <jats:italic>RHD<\/jats:italic> gene. Combination of both methods for fetal cfDNA verification succeeded in 75% of tested group.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Implementation of analysis of three <jats:italic>RHD<\/jats:italic> exons and maternal genomic DNA to routine practice lowers dramatically the ratio of false positive and negative results. This method enables more accurate determination of fetal\u00a0<jats:italic>RHD<\/jats:italic> status with the reduction of unnecessary medical care and RhD immunoprophylaxis.<\/jats:p><\/jats:sec>","DOI":"10.1111\/vox.13099","type":"journal-article","created":{"date-parts":[[2021,3,24]],"date-time":"2021-03-24T20:59:15Z","timestamp":1616619555000},"page":"1012-1019","source":"Crossref","is-referenced-by-count":7,"title":["Optimization of diagnostic strategy for non\u2010invasive cell\u2010free foetal <i>RHD<\/i> determination from maternal plasma"],"prefix":"10.1111","volume":"116","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6117-2127","authenticated-orcid":false,"given":"Eva","family":"Pazourkova","sequence":"first","affiliation":[{"name":"Institute of Biology and Medical Genetics First Faculty of Medicine Charles University and General University Hospital in Prague Praha Czech Republic"},{"name":"Department of Nephrology First Faculty of Medicine Charles University and General University Hospital in Prague Praha Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2891-458X","authenticated-orcid":false,"given":"Iveta","family":"Zednikova","sequence":"additional","affiliation":[{"name":"Institute of Biology and Medical Genetics First Faculty of Medicine Charles University and General University Hospital in Prague Praha Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Korabecna","sequence":"additional","affiliation":[{"name":"Institute of Biology and Medical Genetics First Faculty of Medicine Charles University and General University Hospital in Prague Praha Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jana","family":"Kralova","sequence":"additional","affiliation":[{"name":"Department of Immunohematology Institute of Hematology and Blood Transfusion Prague Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Pisacka","sequence":"additional","affiliation":[{"name":"Department of Immunohematology Institute of Hematology and Blood Transfusion Prague Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michaela","family":"Novotna","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology First Faculty of Medicine Charles University and General University Hospital in Prague Praha Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavel","family":"Calda","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology First Faculty of Medicine Charles University and General University Hospital in Prague Praha Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ales","family":"Horinek","sequence":"additional","affiliation":[{"name":"Institute of Biology and Medical Genetics First Faculty of Medicine Charles University and General University Hospital in Prague Praha Czech Republic"},{"name":"3rd Department of Medicine Department of Endocrinology and Metabolism First Faculty of Medicine Charles University and General University Hospital in Prague Praha Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,3,24]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/vox.12265"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199812103392402"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1423-0410.2004.00569.x"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.siny.2007.12.012"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00439-009-0738-2"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1159\/000345682"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.2001.41010045.x"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1423-0410.1986.tb01958.x"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0030711"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-0528.2006.00988.x"},{"key":"e_1_2_9_12_1","first-page":"CD000020","article-title":"Anti\u2010D administration in pregnancy for preventing Rhesus alloimmunisation","author":"Crowther CA","year":"2013","journal-title":"Cochrane Database Syst Rev"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1054\/blre.1999.0123"},{"key":"e_1_2_9_14_1","first-page":"362","article-title":"Klinick\u00fd v\u00fdznam neinvazivn\u00edho stanoven\u00ed RHD a RHCE genotypu plodu v managementu t\u011bhotenstv\u00ed s rizikem rozvoje hemolytick\u00e9 nemoci plodu a novorozence. 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