{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T18:43:23Z","timestamp":1761677003917},"reference-count":18,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2021,6,21]],"date-time":"2021-06-21T00:00:00Z","timestamp":1624233600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Due to the strong association between ankylosing spondylitis and Human Leukocyte Antigen (HLA)-B27, accurate identification of HLA-B27 is important in the diagnosis of patients with suspected spondyloarthritides. For this study, we compared a high-resolution HLA-B typing method to the clinical flow cytometry and allele-specific PCR melting assays to determine clinical benefits of high-resolution testing.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Residual clinical samples submitted for HLA-B27 testing by flow cytometry were tested by single-locus HLA-B genotyping using next-generation sequencing (NGS), and PCR with melting curve analysis, currently used as a reflex test for indeterminate flow cytometry results.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Fifty out of the 51 samples (98%) positive by flow cytometry confirmed as HLA-B27 positive by PCR melting assay and by NGS. The sample that did not confirm was genotyped as HLA-B*07:02. All the samples negative by flow cytometry were confirmed as HLA-B27 negative by both PCR melting assay and NGS. For the group that was indeterminate by flow cytometry, 84.5% (n\u2009=\u200949) typed as positive for HLA-B27, while 15.5% (n\u2009=\u20099) were negative for HLA-B27 but positive for HLA-B*07:02. NGS was the only method able to distinguish between pathogenic and nonpathogenic HLA-B27 variants, in contrast to the flow cytometry or the PCR melting assays.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>Single-locus NGS is superior to flow cytometry and PCR melting assay for the unambiguous identification of HLA-B27 variants, and uniquely able to distinguish between pathogenic and nonpathogenic B27 alleles. Due to its high accuracy, it may be a feasible superior alternative to flow cytometry and traditional molecular methods for clinical HLA-B27 testing.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/jalm\/jfab046","type":"journal-article","created":{"date-parts":[[2021,5,3]],"date-time":"2021-05-03T19:47:40Z","timestamp":1620071260000},"page":"1221-1227","source":"Crossref","is-referenced-by-count":7,"title":["Comparison of Next-Generation Sequencing-Based Human Leukocyte Antigen Typing with Clinical Flow Cytometry and Allele-Specific PCR Melting Assays for HLA-B27 Genotyping"],"prefix":"10.1093","volume":"6","author":[{"given":"Tracie","family":"Profaizer","sequence":"first","affiliation":[{"name":"ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kimberly","family":"Dibb","sequence":"additional","affiliation":[{"name":"Histocompatibility & Immunogenetics Laboratory, University of Utah Health, Salt Lake City, UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Holly","family":"Bethers","sequence":"additional","affiliation":[{"name":"ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cassandra","family":"Monds","sequence":"additional","affiliation":[{"name":"ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John","family":"Andreasen","sequence":"additional","affiliation":[{"name":"ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julio C","family":"Delgado","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eszter","family":"L\u00e1z\u00e1r-Moln\u00e1r","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2021,6,21]]},"reference":[{"key":"2021090112213158400_jfab046-B1","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1097\/MAJ.0b013e31820f8c99","article-title":"Epidemiology of spondyloarthritis in North America","volume":"341","author":"Reveille","year":"2011","journal-title":"Am J Med Sci"},{"key":"2021090112213158400_jfab046-B2","doi-asserted-by":"crossref","first-page":"904","DOI":"10.1016\/S0140-6736(73)91360-3","article-title":"Ankylosing spondylitis and HL-A 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