{
  "abstract": "Background\n\nLow-frequency heteroplasmic mitochondrial DNA (mtDNA) variants are associated with aging and neurological diseases, including Parkinson's disease (PD). Targeted deep mtDNA sequencing using PacBio HiFi long reads has the potential to resolve heteroplasmy across the full mitochondrial genome with high accuracy.Methods\n\nTo validate Vega PacBio sequencing for detecting mtDNA heteroplasmy, we analyzed four predefined mixtures of two mtDNA haplotypes. We generated a single long-range PCR amplicon covering the entire mitochondrial genome. These amplicons were mixed at predefined ratios (minor mixture haplotype component: 5%, 2%, 1%, and 0.1%). Variant calling was performed using Mutserve2, and accuracy was assessed by calculating the F₁ score from comparisons between expected and detected variants. Full-length mtDNA PacBio sequencing was applied to explore heteroplasmy across fibroblast passages derived from five LRRK2 p.Gly2019Ser variant carriers (n = 3 affected with PD and n = 2 unaffected carriers). Changes in mtDNA heteroplasmy level and variant load were assessed longitudinally using a linear mixed model as a proof-of-principle.Results\n\nThe single-amplicon approach enabled full-length haplotype resolution without amplification bias associated with overlapping PCR strategies. The F₁ score of the predefined mixtures was 1.0 for heteroplasmy levels between 5% and 1% and remained high (0.91) at 0.1%. At the specifically lowered calling threshold applied to the 0.1% mixture, n = 10/62 additional variants discordant with the Illumina reference were observed, but sensitivity remained very high at 1.00 in that mixture. Detected minor variants closely matched expected heteroplasmy levels, with average variant levels of 0.057 (5%), 0.022 (2%), 0.011 (1%), and 0.001 (0.1%). Across twelve fibroblast passages, we observed fewer mtDNA heteroplasmic variants. In this small proof-of-principle experiment, heteroplasmic variant load was higher in affected (n = 3) than in unaffected (n = 2) LRRK2 variant carriers and with older age. Notably, we observed distinct patterns of heteroplasmic variants that either increased or decreased in heteroplasmy level across passages.Conclusion\n\nPacBio HiFi sequencing, combined with a single-amplicon strategy, enables accurate full-length mtDNA heteroplasmy detection and longitudinal analysis, providing a valuable tool for studying mitochondrial variation and dynamics in disease.",
  "abstract_shown": true,
  "affiliations": [
    "Institute of Neurogenetics, University of Lübeck, Lübeck, Germany.",
    "Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg.",
    "Institute of Genetic Epidemiology, Medical University of Innsbruck, Innsbruck, Austria.",
    "Institute of Neurogenetics, University of Lübeck, Lübeck, Germany. joanne.trinh@uni-luebeck.de."
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 10,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Theresa Lüth"
    },
    {
      "affs": [
        0
      ],
      "name": "Susen Schaake"
    },
    {
      "affs": [
        0
      ],
      "name": "Christoph Much"
    },
    {
      "affs": [
        0
      ],
      "name": "Mary Megan Belyea"
    },
    {
      "affs": [
        0
      ],
      "name": "Philip Seibler"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Anne Grünewald"
    },
    {
      "affs": [
        1
      ],
      "name": "Patrick May",
      "orcid": "0000-0001-8698-3770"
    },
    {
      "affs": [
        0
      ],
      "name": "Christine Klein"
    },
    {
      "affs": [
        2
      ],
      "name": "Hansi Weissensteiner"
    },
    {
      "affs": [
        3
      ],
      "name": "Joanne Trinh"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/3g6ae6snvckf3.bib",
  "cited_by": 0,
  "doi": "10.1186/s12864-026-13426-y",
  "fulltext_url": "https://europepmc.org/articles/PMC13651137",
  "id": "3g6ae6snvckf3",
  "issns": [
    "1471-2164"
  ],
  "landing_url": "https://europepmc.org/article/MED/42850586",
  "language": "eng",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|Mitochondrial Function and Pathology"
    ]
  ],
  "openalex": "W7221031662",
  "pdf_url": "https://europepmc.org/articles/PMC13651137?pdf=render",
  "pmcid": "PMC13651137",
  "pmid": "42850586",
  "published": "2026-10-08",
  "publisher": "Springer Science and Business Media LLC",
  "ris": "https://research.draconitic.com/paper/3g6ae6snvckf3.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:18:02Z",
      "native_id": "MED:42850586",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42850586"
    },
    {
      "first_seen": "2026-10-30T12:19:08Z",
      "native_id": "10.1186/s12864-026-13426-y",
      "source": "crossref",
      "url": "https://link.springer.com/10.1186/s12864-026-13426-y"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Mitochondrial Function and Pathology",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Long-read sequencing enables high-accuracy mitochondrial heteroplasmy detection in Parkinson's disease.",
  "title_text": "Long-read sequencing enables high-accuracy mitochondrial heteroplasmy detection in Parkinson's disease.",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:19:08Z",
  "url": "https://research.draconitic.com/paper/3g6ae6snvckf3",
  "venue": "BMC genomics",
  "versions": []
}