{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:49:27Z","timestamp":1787028567893,"version":"build-2736575974"},"reference-count":31,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2025,8,10]],"date-time":"2025-08-10T00:00:00Z","timestamp":1754784000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Monoquant P\/L."}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The role of HAT-PCR (High A\/T or High Annealing Temperature\u2013PCR) in the quantification of minimal residual disease (MRD) was investigated in chronic lymphocytic leukemia (CLL) and myeloma. The IGH gene sequence was determined by next-generation sequencing (NGS), either by the Lymphotrack kit or by preparing libraries using an in-house two-round PCR protocol which enabled successful sequencing in 37\/37 CLL marrow samples and 34\/35 myeloma marrow samples. MRD was quantified by HAT-PCR in 125 CLL marrow or blood samples from 36 patients, with 2 results being less than 10\u22126 and in 63 myeloma marrow samples from 35 patients, with 10 results being less than 10\u22126. Measurement of MRD in 113 pairs of CLL samples and 51 pairs of myeloma samples showed that HAT-PCR was significantly more sensitive than flow. Compared to marrow MRD, blood MRD was relatively high in CLL but very low or undetectable in myeloma. Flow-positive HAT-PCR negative samples were not seen in myeloma, although the literature review suggested that flow-positive NGS-negative myeloma samples are sometimes observed. The ability of HAT-PCR to quantify down to and below 10\u22126 and the practical advantages of PCR suggest that HAT-PCR could be used widely for the quantification of MRD in lymphoid malignancy.<\/jats:p>","DOI":"10.3390\/ijms26167720","type":"journal-article","created":{"date-parts":[[2025,8,11]],"date-time":"2025-08-11T11:02:09Z","timestamp":1754910129000},"page":"7720","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["HAT-PCR Enables Sensitive Quantification of Minimal Residual Disease in Chronic Lymphocytic Leukemia and Myeloma"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-3767-2758","authenticated-orcid":false,"given":"Elizabeth","family":"Hughes","sequence":"first","affiliation":[{"name":"School of Medicine, Flinders University, Adelaide 5042, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sue","family":"Latham","sequence":"additional","affiliation":[{"name":"School of Medicine, Flinders University, Adelaide 5042, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5811-1331","authenticated-orcid":false,"given":"Bryone","family":"Kuss","sequence":"additional","affiliation":[{"name":"School of Medicine, Flinders University, Adelaide 5042, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott","family":"Grist","sequence":"additional","affiliation":[{"name":"Genomix, Adelaide 5042, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel","family":"Hall","sequence":"additional","affiliation":[{"name":"Genomix, Adelaide 5042, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9376-4223","authenticated-orcid":false,"given":"Tiffany","family":"Khong","sequence":"additional","affiliation":[{"name":"Alfred Hospital, Melbourne 3181, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malgorzata","family":"Gorniak","sequence":"additional","affiliation":[{"name":"Alfred Hospital, Melbourne 3181, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"Spencer","sequence":"additional","affiliation":[{"name":"Alfred Hospital, Melbourne 3181, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Constantine","family":"Tam","sequence":"additional","affiliation":[{"name":"Alfred Hospital, Melbourne 3181, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen","family":"Mulligan","sequence":"additional","affiliation":[{"name":"School of Medicine, University of 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