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Analyzing the public database, we identified the methylated promoter regions of the genes <jats:italic>FGD5<\/jats:italic>, <jats:italic>GPC6<\/jats:italic>, and <jats:italic>MSC<\/jats:italic>. Using digital polymerase chain reaction (dPCR), we termed the \u201camplicon of methylated sites using a specific enzyme\u201d assay as \u201cAMUSE.\u201d We examined 180 and 114 pre\u2010 and postoperative serial plasma samples from 28 recurrent and 19 recurrence\u2010free pathological stage III CRC patients, respectively. The results showed 22 AMUSE\u2010positive of 28 recurrent patients (sensitivity, 78.6%) and 17 AMUSE\u2010negative of 19 recurrence\u2010free patients (specificity, 89.5%). AMUSE predicted recurrence 208\u2009days before conventional diagnosis using radiological imaging. Regarding ACT evaluation by the reactive response, 19 AMUSE\u2010positive patients during their second or third blood samples showed a significantly poorer prognosis than the other patients (<jats:italic>p<\/jats:italic>\u2009=\u20099E\u201004). The AMUSE assay stratified four groups by the altered patterns of tumor burden postoperatively. Interestingly, only 34.8% of cases tested AMUSE\u2010negative during ACT treatment, indicating eligibility for ACT. The AMUSE assay addresses the clinical need for accurate MRD monitoring with universal applicability, minimal invasiveness, and cost\u2010effectiveness, thereby enabling the timely detection of recurrences. This assay can effectively evaluate the efficacy of ACT in patients with stage III CRC following curative resection. Our study strongly recommends reevaluating the clinical application of ACT using the AMUSE assay.<\/jats:p>","DOI":"10.1111\/cas.16149","type":"journal-article","created":{"date-parts":[[2024,3,27]],"date-time":"2024-03-27T03:45:25Z","timestamp":1711511125000},"page":"1989-2001","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Implementable assay for monitoring minimum residual disease after radical treatment for colorectal cancer"],"prefix":"10.1111","volume":"115","author":[{"given":"Takafumi","family":"Nakano","sequence":"first","affiliation":[{"name":"Department of Surgery Kyushu University Beppu Hospital  Beppu Japan"},{"name":"Department of Surgery The Jikei University School of Medicine  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seiichiro","family":"Takao","sequence":"additional","affiliation":[{"name":"Department of Surgery Kyushu University Beppu Hospital  Beppu 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