{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T00:52:36Z","timestamp":1774659156354,"version":"3.50.1"},"reference-count":38,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2019,12,1]],"date-time":"2019-12-01T00:00:00Z","timestamp":1575158400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["HLA"],"published-print":{"date-parts":[[2020,3]]},"abstract":"<jats:p>The linkage between the occurrence of human leucocyte antigen <jats:italic>B*27<\/jats:italic> (<jats:italic>HLA\u2010B*27<\/jats:italic>) and ankylosing spondylitis or other related spondyloarthritides is well documented. PCR based methods are widely used for <jats:italic>HLA\u2010B*27<\/jats:italic> screening. To refine <jats:italic>HLA\u2010B*27<\/jats:italic> testing we aimed at establishing a real\u2010time PCR protocol to detect the <jats:italic>HLA\u2010B*27<\/jats:italic> allele directly in blood samples, without DNA extraction. <jats:italic>HLA\u2010B*27<\/jats:italic> analysis was performed by two real\u2010time PCRs using TaqMan primer\u2010probe assays for <jats:italic>B*27<\/jats:italic> specific amplification of exon 2 or exon 3 of the HLA\u2010B gene together with a mutant of Taq polymerase for direct blood PCR. Conditions for direct blood PCR were optimized and the reliability of the direct blood PCR protocol was evaluated by re\u2010genotyping over 200 blood samples from patients who previously underwent routine DNA\u2010based <jats:italic>HLA\u2010B*27<\/jats:italic> testing. Heating blood samples at 95\u00b0C for 10\u2009minutes significantly improved PCR performance. Results from real\u2010time PCR based <jats:italic>HLA\u2010B*27<\/jats:italic> testing directly in blood of over 200 patients were in 100% concordance with results obtained by routine DNA\u2010based <jats:italic>HLA\u2010B*27<\/jats:italic> genotyping. In summary, we present a reliable real\u2010time PCR protocol for <jats:italic>HLA\u2010B*27<\/jats:italic> screening directly in whole blood supporting fast clarification of the presence of ankylosing spondylitis or other spondyloarthritides in suspected cases.<\/jats:p>","DOI":"10.1111\/tan.13767","type":"journal-article","created":{"date-parts":[[2019,11,21]],"date-time":"2019-11-21T02:10:01Z","timestamp":1574302201000},"page":"189-195","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Real\u2010time PCR based <i>HLA\u2010B*27<\/i> screening directly in whole blood"],"prefix":"10.1111","volume":"95","author":[{"given":"Kathrin","family":"Geiger","sequence":"first","affiliation":[{"name":"Vorarlberg Institute for Vascular Investigation and Treatment (VIVIT)  Feldkirch 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