{
  "abstract": "DNA methylation provides informative signals for liquid biopsy, but low template abundance, distributed signals across loci and conversion-dependent processing can complicate implementation in low-input samples. Here we present Delta-HLD, a PCR-compatible platform that quantifies methylation directly in native DNA through sequential hybridization, ligation and methylation-sensitive digestion. By encoding locus specificity at the hemi-probe hybridization and ligation stage, Delta-HLD allows methylation-dependent products from multiple loci to converge on a shared amplification and detection system and contribute to a single panel-level readout. We established the assay chemistry, optimized panel composition through model-guided experiments and implemented the platform in qPCR and ddPCR, including a triplex qPCR format with per-sample digestion and amplification controls. Proof-of-concept plasma analyses showed discriminatory signals in colorectal cancer and supported transfer of the same assay logic to hepatocellular carcinoma. The assay was further adapted for cfDNA analysis following platelet-retaining processing, preserving cfDNA-associated signal while reducing genomic DNA recognition. Delta-HLD provides a compact native-DNA platform for multilocus methylation analysis in low-input settings without base conversion.",
  "abstract_shown": true,
  "affiliations": [
    "Instituto de Histologia y Embriologia de Mendoza (IHEM)-CONICET, Universidad Nacional de Cuyo (UNCUYO), Mendoza, Argentina; Facultad de Ciencias Medicas, Univer"
  ],
  "article_type": "PUBLISHAHEADOFPRINT",
  "arxiv": null,
  "author_total": 24,
  "authors": [
    {
      "name": "C. C. Vaquer"
    },
    {
      "name": "P. A. Wetten"
    },
    {
      "name": "C. Garcia Samartino"
    },
    {
      "name": "J. D. Rodriguez"
    },
    {
      "name": "N. R. Manzino"
    },
    {
      "name": "R. Perez Ravier"
    },
    {
      "name": "A. R. Angeloni"
    },
    {
      "name": "R. D. Militello"
    },
    {
      "name": "A. Ledesma"
    },
    {
      "name": "J. Sesma"
    },
    {
      "name": "Y. Agnella"
    },
    {
      "name": "E. Gudino"
    },
    {
      "name": "C. E. Fracchia Diaz"
    },
    {
      "name": "R. Ongay"
    },
    {
      "name": "G. Sanguinetti"
    },
    {
      "name": "A. Correa"
    },
    {
      "name": "M. Carlen"
    },
    {
      "name": "W. R. Minatti"
    },
    {
      "name": "G. A. Vaschalde"
    },
    {
      "name": "P. Valdemoros"
    },
    {
      "name": "L. Sarrio"
    },
    {
      "name": "L. Mayorga"
    },
    {
      "name": "V. Bocanegra"
    },
    {
      "affs": [
        0
      ],
      "name": "E. M. Campoy"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/bdsbwahad0wzr.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.06.11.26354897",
  "fulltext_url": "https://www.medrxiv.org/content/10.64898/2026.06.11.26354897v2.full",
  "id": "bdsbwahad0wzr",
  "issns": [],
  "landing_url": "https://www.medrxiv.org/content/10.64898/2026.06.11.26354897v2",
  "language": null,
  "licence": "CC-BY-NC-ND",
  "licence_label": "CC BY-NC-ND",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "medrxiv",
      "oncology"
    ],
    [
      "openalex",
      "1312|Epigenetics and DNA Methylation"
    ]
  ],
  "openalex": "W7165515610",
  "pdf_url": "https://www.medrxiv.org/content/10.64898/2026.06.11.26354897v2.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-06-11",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/bdsbwahad0wzr.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:20:34Z",
      "native_id": "10.64898/2026.06.11.26354897",
      "source": "medrxiv",
      "url": "https://www.medrxiv.org/content/10.64898/2026.06.11.26354897v2"
    }
  ],
  "subjects": [
    {
      "field": "medicine",
      "origin": "medrxiv:oncology",
      "subfield": "oncology"
    }
  ],
  "title": "Panel-level multilocus methylation quantification in native cell-free DNA by PCR-compatible sequential enzymatic processing",
  "title_text": "Panel-level multilocus methylation quantification in native cell-free DNA by PCR-compatible sequential enzymatic processing",
  "type": "preprint",
  "updated_at": "2026-10-30T12:20:34Z",
  "url": "https://research.draconitic.com/paper/bdsbwahad0wzr",
  "venue": "medRxiv",
  "versions": []
}