{
  "abstract": "Background: Next-generation sequencing (NGS) is essential for precision oncology, yet its clinical implementation is often hindered by limited sample quantity, complex workflows, and cross-contamination risks. We evaluated a modified One-Step Multiplex PCR (OS-MPCR) assay designed to streamline library construction while maintaining high analytical sensitivity and diagnostic accuracy. Methods: The OS-MPCR method integrates adapter ligation and library amplification into a single, closed-tube reaction, eliminating intermediate open-tube manipulations. The analytical performance was evaluated using standard reference materials targeting SNVs, Indels, and fusions. Clinical validation was performed on retrospective formalin-fixed paraffin-embedded (FFPE) specimens (DNA, N = 76; RNA, N = 34) from lung cancer patients, using well-established liquid-phase hybrid capture assays as the clinical verification standard. Results: OS-MPCR streamlined the workflow, reducing hands-on time to just 20 minutes. The assay enabled reliable variant detection at 1% frequency with as little as 1 ng of DNA for SNVs/Indels and 5 ng of total RNA for fusions, with low-input quantification limits inherently bounded by absolute molecular copy numbers. In the clinical cohort, OS-MPCR demonstrated 100% diagnostic concordance with the verification method. Quantitative analysis revealed a strong correlation in variant allele frequencies (Spearman r = 0.9777, 95% CI: 0.9668–0.9851, P < 0.0001), with libraries consistently achieving highly reproducible yields and stable, near 1.0 target rates across the entire cohort. Conclusion: OS-MPCR offers a streamlined and low-input alternative to conventional multi-step multiplex PCR methods. By combining reduced hands-on time and intrinsic contamination control with high analytical accuracy, this approach represents a practical refinement for targeted amplicon sequencing in diagnostic workflows.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Pathology, Zibo First Hospital, Zibo, Shandong, China",
    "Shanghai Yijian Health Technology Co., Ltd., Shanghai, China"
  ],
  "article_type": "Research Article",
  "arxiv": null,
  "author_total": 9,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Haiyan Feng"
    },
    {
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        1
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      "name": "Xiao Hu"
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    {
      "name": "Lei Zhang"
    },
    {
      "name": "Qi Zhang"
    },
    {
      "name": "Shuocun Wang"
    },
    {
      "name": "Bingqing Lv"
    },
    {
      "name": "Yufeng Zhang"
    },
    {
      "name": "Dong Shi"
    },
    {
      "name": "Ping Zhou"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/fv9gjysxd85px.bib",
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  "doi": "10.1371/journal.pone.0357776",
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  "id": "fv9gjysxd85px",
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    "1932-6203"
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  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1306|Cancer Genomics and Diagnostics"
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    [
      "plos",
      "/Biology and life sciences/Biochemistry/Nucleic acids/DNA/DNA libraries"
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      "/Biology and life sciences/Genetics/DNA/DNA libraries"
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      "plos",
      "/Biology and life sciences/Genetics/Genomics"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/Artificial gene amplification and extension/Polymerase chain reaction"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/Molecular biology assays and analysis techniques/Nucleic acid analysis/RNA analysis"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/Molecular probe techniques/Probe hybridization/DNA-RNA hybridization"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/Sequencing techniques/RNA sequencing"
    ],
    [
      "plos",
      "/Medicine and health sciences/Oncology"
    ],
    [
      "plos",
      "/Medicine and health sciences/Pulmonology/Pleural effusion"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/Artificial gene amplification and extension/Polymerase chain reaction"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/Molecular biology assays and analysis techniques/Nucleic acid analysis/RNA analysis"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/Molecular probe techniques/Probe hybridization/DNA-RNA hybridization"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/Sequencing techniques/RNA sequencing"
    ]
  ],
  "openalex": "W7213306582",
  "pdf_url": "https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0357776&type=printable",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-15",
  "publisher": "Public Library of Science",
  "ris": "https://research.draconitic.com/paper/fv9gjysxd85px.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:49Z",
      "native_id": "10.1371/journal.pone.0357776",
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      "url": "https://doi.org/10.1371/journal.pone.0357776"
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  "subjects": [
    {
      "field": "biochemistry",
      "origin": "plos:molecular biology",
      "subfield": "molecular-biology"
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    {
      "field": "biology",
      "origin": "plos:genetics",
      "subfield": "genetics-genomics"
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  ],
  "title": "A novel one-step multiplex PCR method (OS-MPCR) for efficient NGS library preparation in cancer",
  "title_text": "A novel one-step multiplex PCR method (OS-MPCR) for efficient NGS library preparation in cancer",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:17:49Z",
  "url": "https://research.draconitic.com/paper/fv9gjysxd85px",
  "venue": "PLOS One",
  "versions": []
}