{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T11:31:30Z","timestamp":1771500690821,"version":"3.50.1"},"reference-count":15,"publisher":"BMJ","issue":"3","content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Clin Pathol"],"accepted":{"date-parts":[[2025,12,27]]},"published-print":{"date-parts":[[2026,3]]},"abstract":"<jats:sec>\n                    <jats:title>Aims<\/jats:title>\n                    <jats:p>\n                      The diagnosis and therapeutic decision-making in chronic myeloid leukaemia (CML) depend on the detection and quantification of\n                      <jats:italic>BCR::ABL1<\/jats:italic>\n                      . Currently, no clinical assay is used in routine practice to simultaneously quantify multiple\n                      <jats:italic>BCR::ABL1<\/jats:italic>\n                      isoforms, requiring separate tests for each isoform. To address this challenge, we have developed and optimised an assay (BloodHound) that leverages quantitative reverse transcription PCR (RT-qPCR) and high-resolution melting (HRM) technologies to simultaneously detect and quantify four\n                      <jats:italic>BCR::ABL1<\/jats:italic>\n                      isoforms (p190, p210, p230 and p203), with a limit of detection of 0.001%.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      To evaluate the clinical utility of this assay, we analysed 895 peripheral blood and bone marrow samples from patients with suspected, established or relapsed CML. Sanger sequencing was used as an orthogonal method to confirm fusion junction identity, and the Qiagen ipsogen\n                      <jats:italic>BCR::ABL1<\/jats:italic>\n                      RT-qPCR assay was used for quantitative comparison.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      <jats:italic>BCR::ABL1<\/jats:italic>\n                      was detected in 20.9% of samples, with p210 alone being the most prevalent (86.1% (161\/187), 95\u2009CI 80.3% to 90.7%), followed by coexpression of p190 and p210 (12.3% (23\/187), 95%\u2009CI 7.6% to 17.0%), while p190 alone (1.1% (2\/187), 95%\u2009CI 0.1% to 3.8%) and p230 alone (0.5% (1\/187), 95%\u2009CI 0% to 2.9%) were rare and p203 was not detected. The assay demonstrated high sensitivity and specificity, with 100% concordance with Sanger sequencing and showed excellent agreement with the Qiagen\n                      <jats:italic>BCR::ABL1<\/jats:italic>\n                      RT-qPCR assay (r=0.998).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>The present multiplex RT-qPCR\/HRM assay may help streamline clinical workflows, enhance diagnostic precision and offer a practical platform for studying CML clonal dynamics. It also holds promise for facilitating interlaboratory standardisation of non-p210 quantification.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/jcp-2025-210447","type":"journal-article","created":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T18:20:20Z","timestamp":1767896420000},"page":"192-197","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":0,"title":["Quantitative reverse transcription PCR coupled with high-resolution melting for simultaneous detection and quantification of four isoforms of BCR::ABL1"],"prefix":"10.1136","volume":"79","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7405-1845","authenticated-orcid":false,"given":"Ting","family":"Zhou","sequence":"first","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ayman","family":"Mohamed","sequence":"additional","affiliation":[{"name":"Precipio, New Haven, Connecticut, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ben L","family":"Legendre","sequence":"additional","affiliation":[{"name":"Precipio, New Haven, Connecticut, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heather","family":"Newell","sequence":"additional","affiliation":[{"name":"Precipio, New Haven, Connecticut, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kojo S J","family":"Elenitoba-Johnson","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"239","published-online":{"date-parts":[[2026,1,8]]},"reference":[{"key":"2026021902550847000_79.3.192.1","doi-asserted-by":"publisher","DOI":"10.1038\/243290a0"},{"key":"2026021902550847000_79.3.192.2","doi-asserted-by":"publisher","DOI":"10.1007\/s11899-018-0474-6"},{"key":"2026021902550847000_79.3.192.3","doi-asserted-by":"crossref","first-page":"5213","DOI":"10.1182\/blood.V87.12.5213.bloodjournal87125213","article-title":"p190 BCR-ABL mRNA is expressed at low levels in p210-positive chronic myeloid and acute lymphoblastic leukemias","volume":"87","author":"van Rhee","year":"1996","journal-title":"Blood"},{"key":"2026021902550847000_79.3.192.4","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.trsl.2006.07.002","article-title":"Comprehensive analysis of BCR-ABL transcript types in Korean CML patients using a newly developed multiplex RT-PCR","volume":"148","author":"Goh","year":"2006","journal-title":"Transl Res"},{"key":"2026021902550847000_79.3.192.5","doi-asserted-by":"crossref","DOI":"10.1038\/bcj.2017.62","article-title":"Clinical and prognostic significance of e1a2 BCR-ABL1 transcript subtype in chronic myeloid leukemia","volume":"7","author":"Gong","year":"2017","journal-title":"Blood Cancer J"},{"key":"2026021902550847000_79.3.192.6","doi-asserted-by":"publisher","DOI":"10.1038\/s41375-020-01082-4"},{"key":"2026021902550847000_79.3.192.7","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1111\/ijlh.12616","article-title":"Frequency of rare BCR-ABL1 fusion transcripts in chronic myeloid leukemia patients","volume":"39","author":"Arun","year":"2017","journal-title":"Int J Lab Hematol"},{"key":"2026021902550847000_79.3.192.8","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1038\/s41375-018-0341-4","article-title":"The proportion of different BCR-ABL1 transcript types in chronic myeloid leukemia. An international overview","volume":"33","author":"Baccarani","year":"2019","journal-title":"Leukemia"},{"key":"2026021902550847000_79.3.192.9","doi-asserted-by":"crossref","first-page":"2041","DOI":"10.1038\/s41375-024-02314-7","article-title":"The e13a3 (b2a3) and e14a3 (b3a3) BCR::ABL1 isoforms are resistant to asciminib","volume":"38","author":"Leske","year":"2024","journal-title":"Leukemia"},{"key":"2026021902550847000_79.3.192.10","doi-asserted-by":"publisher","DOI":"10.2353\/jmoldx.2009.090008"},{"key":"2026021902550847000_79.3.192.11","doi-asserted-by":"publisher","DOI":"10.3109\/10428194.2015.1037751"},{"key":"2026021902550847000_79.3.192.12","first-page":"191","article-title":"Appearance of acute leukemia-associated P190BCR-ABL in chronic myelogenous leukemia may correlate with disease progression","volume":"5","author":"Dhingra","year":"1991","journal-title":"Leukemia"},{"key":"2026021902550847000_79.3.192.13","doi-asserted-by":"crossref","first-page":"1707","DOI":"10.1159\/000520400","article-title":"Navigating Challenges in Monitoring Chronic Myeloid Leukemia with Multiple BCR-ABL1 Transcripts","volume":"14","author":"Smith","year":"2021","journal-title":"Case Rep Oncol"},{"key":"2026021902550847000_79.3.192.14","doi-asserted-by":"crossref","DOI":"10.1186\/s40064-015-0930-x","article-title":"Coexistence of P190 and P210 BCR\/ABL transcripts in chronic myeloid leukemia blast crisis resistant to imatinib","volume":"4","author":"Junmei","year":"2015","journal-title":"Springerplus"},{"key":"2026021902550847000_79.3.192.15","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2012.196477"}],"container-title":["Journal of Clinical Pathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1136\/jcp-2025-210447","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T10:55:40Z","timestamp":1771498540000},"score":1,"resource":{"primary":{"URL":"https:\/\/jcp.bmj.com\/lookup\/doi\/10.1136\/jcp-2025-210447"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,8]]},"references-count":15,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,2,19]]},"published-print":{"date-parts":[[2026,3]]}},"alternative-id":["10.1136\/jcp-2025-210447"],"URL":"https:\/\/doi.org\/10.1136\/jcp-2025-210447","relation":{},"ISSN":["0021-9746","1472-4146"],"issn-type":[{"value":"0021-9746","type":"print"},{"value":"1472-4146","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,8]]}}}