{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T13:00:08Z","timestamp":1790686808541,"version":"4.1.0"},"reference-count":28,"publisher":"Wiley","issue":"18","license":[{"start":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T00:00:00Z","timestamp":1786579200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T00:00:00Z","timestamp":1786579200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100005089","name":"Natural Science Foundation of Beijing Municipality","doi-asserted-by":"publisher","award":["L244070"],"award-info":[{"award-number":["L244070"]}],"id":[{"id":"10.13039\/501100005089","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Clinical Laboratory Analysis"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Immune\u2010mediated platelet transfusion refractoriness (IPTR) remains a major clinical challenge, with immune alloimmunization against HLA and HPA as the predominant causes. Although HLA cross\u2010reactive groups (CREGs) provide a practical alternative to allele\u2010level matching, methods for simultaneous HLA\u2010CREG and HPA genotyping remain limited despite their potential to improve platelet matching.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Specific U\u2010anchoring primers (UAPs) and amplification refractory mutation system (ARMS) primers were designed and optimized based on one or more SNPs within HLA\u2010CREG subgroups and HPA alleles. Reaction conditions were optimized, and analytical performance was evaluated. A total of 200 donor DNA samples were tested and compared with NGS and a commercial HPA sequencing kit, with discrepant results verified by Sanger sequencing.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We developed and optimized a novel multiplex real\u2010time quantitative PCR (qPCR) assay integrating UAPs and ARMS primers for simultaneous detection of HLA\u2010A\/B CREGs and major HPA alleles in 12 reaction tubes. The assay demonstrated an overall limit of detection (LOD) of 0.838\u2009ng\/\u03bcL and the intra\u2010\/inter\u2010assay coefficients of variation were both below 5%, indicating high reproducibility. HLA\u2010CREG typing results were 100% concordant with next\u2010generation sequencing. Sixteen discrepant HPA genotyping results were identified compared with the commercial kit, all of which were confirmed by Sanger sequencing and were consistent with the multiplex qPCR assay.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This multiplex qPCR assay enables rapid, accurate, and cost\u2010effective simultaneous detection of HLA\u2010CREGs and HPA alleles, providing a practical tool for comprehensive platelet compatibility assessment and improved transfusion matching.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/jcla.70326","type":"journal-article","created":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T10:39:21Z","timestamp":1786617561000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Practical and Efficient Multiplex\n                    <scp>qPCR<\/scp>\n                    Assay for Simultaneous Detection of\n                    <scp>HLA<\/scp>\n                    Cross\u2010Reactive Groups and Human Platelet Antigens"],"prefix":"10.1002","volume":"40","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7790-1550","authenticated-orcid":false,"given":"Huimin","family":"Ji","sequence":"first","affiliation":[{"name":"National Center for Clinical Laboratories Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Sciences  Beijing People's Republic of China"},{"name":"Beijing Engineering Research Center of Laboratory Medicine  Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaihao","family":"Feng","sequence":"additional","affiliation":[{"name":"National Center for Clinical Laboratories Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Sciences  Beijing People's Republic of China"},{"name":"Beijing Engineering Research Center of Laboratory Medicine  Beijing People's Republic of China"},{"name":"National Center for Clinical Laboratories Peking Union Medical College, Chinese Academy of Medical Sciences  Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4546-2191","authenticated-orcid":false,"given":"Yunping","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Laboratory Shenzhen Blood Center  Shenzhen People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6154-7304","authenticated-orcid":false,"given":"Le","family":"Chang","sequence":"additional","affiliation":[{"name":"National Center for Clinical Laboratories Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Sciences  Beijing People's Republic of China"},{"name":"Beijing Engineering Research Center of Laboratory Medicine  Beijing People's Republic of China"},{"name":"National Center for Clinical Laboratories Peking Union Medical College, Chinese Academy of Medical Sciences  Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8556-7417","authenticated-orcid":false,"given":"Ying","family":"Yan","sequence":"additional","affiliation":[{"name":"National Center for Clinical Laboratories Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Sciences  Beijing People's Republic of China"},{"name":"Beijing Engineering Research Center of Laboratory Medicine  Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2938-818X","authenticated-orcid":false,"given":"Jie","family":"Ma","sequence":"additional","affiliation":[{"name":"National Center for Clinical Laboratories Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Sciences  Beijing People's Republic of China"},{"name":"National Center for Clinical Laboratories Peking Union Medical College, Chinese Academy of Medical Sciences  Beijing People's Republic of China"},{"name":"Center of Biotherapy Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences  Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Liu","sequence":"additional","affiliation":[{"name":"63750 Military Hospital of the People's Liberation Army  Shaanxi People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinfeng","family":"Zeng","sequence":"additional","affiliation":[{"name":"Department of Laboratory Shenzhen Blood Center  Shenzhen People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huizhen","family":"Sun","sequence":"additional","affiliation":[{"name":"National Center for Clinical Laboratories Beijing Hospital, National Center of Gerontology; 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