{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T14:08:38Z","timestamp":1789913318627,"version":"4.0.1"},"reference-count":32,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2012,4,20]],"date-time":"2012-04-20T00:00:00Z","timestamp":1334880000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Prenatal Diagnosis"],"published-print":{"date-parts":[[2012,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Objective<\/jats:title><jats:p>The aim of this study was to develop a pyrophosphorolysis\u2010activated polymerization (PAP) assay for non\u2010invasive prenatal diagnosis (NIPD) of <jats:italic>\u03b2<\/jats:italic>\u2010thalassemia major and sickle\u2010cell disease (SCD). PAP is able to detect mutations in free fetal DNA in a highly contaminating environment of maternal plasma DNA.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Pyrophosphorolysis\u2010activated polymerization primers were designed for 12 informative SNPs, genotyped by melting curve analysis (MCA) in both parents. The PAP assay was tested in a series of 13 plasma DNA samples collected from pregnant women. A retrospective NIPD was performed in a couple at risk for SCD.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>All PAP reactions were optimized and able to detect &lt;3% target gDNA in a background of &gt;97% wildtype gDNA. In all 13 cases, the paternal allele was detected by PAP in maternal plasma at 10 to 18\u2009weeks of gestation. For the couple at risk, PAP showed presence of the normal paternal SNP allele in maternal plasma, which was confirmed by results of the chorionic villus sampling analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>In contrast to other methods used for NIPD, the combined PAP and MCA analysis detecting the normal paternal allele is also applicable for couples at risk carrying the same mutation, provided that a previously born child is available for testing to determine the linkage to the paternal SNPs. \u00a9 2012 John Wiley &amp; Sons, Ltd.<\/jats:p><\/jats:sec>","DOI":"10.1002\/pd.3864","type":"journal-article","created":{"date-parts":[[2012,4,20]],"date-time":"2012-04-20T06:04:25Z","timestamp":1334901865000},"page":"578-587","source":"Crossref","is-referenced-by-count":39,"title":["Non\u2010invasive prenatal diagnosis of beta\u2010thalassemia and sickle\u2010cell disease using pyrophosphorolysis\u2010activated polymerization and melting curve analysis"],"prefix":"10.1002","volume":"32","author":[{"given":"Marion","family":"Phylipsen","sequence":"first","affiliation":[{"name":"Hemoglobinopathies Laboratory, Center for Human and Clinical Genetics Leiden University Medical Center  Leiden The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Supawadee","family":"Yamsri","sequence":"additional","affiliation":[{"name":"Center for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences Khon Kaen University  Khon Kaen Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmely E.","family":"Treffers","sequence":"additional","affiliation":[{"name":"Hemoglobinopathies Laboratory, Center for Human and Clinical Genetics Leiden University Medical Center  Leiden The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diahann T. 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