{
  "abstract": "Pathogens causing respiratory infections have reemerged globally since the COVID-19 pandemic. Rapid and accurate identification of respiratory pathogens is critical for diagnosis and treatment. In this study, we developed and evaluated a multiplex real-time PCR assay with a fluorescence probe melting curve (mqPCR-PMC) assay to simultaneously detect six common pathogens, namely, influenza virus A and B (IFV-A and -B), respiratory syncytial virus (RSV), human rhinovirus (HRV), human adenovirus (HAdV) and Mycoplasma pneumoniae (MP). The primers and probes optimized in this assay exhibited a specificity of 100% with a panel of 57 pathogens. The detection limits of the assay ranged from 248 copies/ml to 394 copies/ml. Moreover, the coefficients of variation ranged from 0.11% to 0.30% for Tm and from 1.59% to 4.10% for Rm. The clinical accuracy of the assay was validated using 760 pharyngeal samples. Compared with a commercial kit based on conventional fluorescence RT-PCR, the assay showed more than 87% sensitivity and 99% specificity for each pathogen. The kappa values ranged from 0.927 to 1.000. Moreover, the assay was more precise for HRV and more sensitive on IFV-A, HAdV, MP and RSV. In summary, the mqPCR-PMC assay was demonstrated to be highly sensitive, highly specific and high throughput, and has potential for clinical use in respiratory infectious disease diagnosis.",
  "abstract_shown": true,
  "affiliations": [
    "Medical Integration and Practice Center, Shandong University, Jinan, Shandong, China",
    "Medical Research and Laboratory Diagnostic Center, Jinan Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China",
    "Research and Development Department, GuoRan Gene Technology (Shandong) Company Limited, Jinan, Shandong, China",
    "Shandong LaiBo Biotechnology Company Limited, Jinan, Shandong, China"
  ],
  "article_type": "Research Article",
  "arxiv": null,
  "author_total": 5,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Mingyu Ji"
    },
    {
      "affs": [
        1
      ],
      "name": "Hongjiao Dong"
    },
    {
      "affs": [
        2
      ],
      "name": "Yihui Xu"
    },
    {
      "affs": [
        3
      ],
      "name": "Yunshan Wang"
    },
    {
      "name": "Qianqian Zhao"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/7fyfmfvckj1z6.bib",
  "cited_by": 0,
  "doi": "10.1371/journal.pone.0357377",
  "fulltext_url": null,
  "id": "7fyfmfvckj1z6",
  "issns": [
    "1932-6203"
  ],
  "landing_url": "https://doi.org/10.1371/journal.pone.0357377",
  "language": null,
  "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",
      "2713|Respiratory viral infections research"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Nucleic acids"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Medical microbiology/Microbial pathogens/Bacterial pathogens"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Medical microbiology/Microbial pathogens/Fungal pathogens"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Medical microbiology/Microbial pathogens/Viral pathogens"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/Artificial gene amplification and extension/Polymerase chain reaction"
    ],
    [
      "plos",
      "/Biology and life sciences/Mycology/Fungal pathogens"
    ],
    [
      "plos",
      "/Biology and life sciences/Organisms/Viruses/Viral pathogens"
    ],
    [
      "plos",
      "/Medicine and health sciences/Medical conditions/Infectious diseases/Respiratory infections"
    ],
    [
      "plos",
      "/Medicine and health sciences/Medical conditions/Respiratory disorders/Respiratory infections"
    ],
    [
      "plos",
      "/Medicine and health sciences/Pathology and laboratory medicine/Pathogens"
    ],
    [
      "plos",
      "/Medicine and health sciences/Pathology and laboratory medicine/Pathogens/Microbial pathogens/Bacterial pathogens"
    ],
    [
      "plos",
      "/Medicine and health sciences/Pathology and laboratory medicine/Pathogens/Microbial pathogens/Fungal pathogens"
    ],
    [
      "plos",
      "/Medicine and health sciences/Pathology and laboratory medicine/Pathogens/Microbial pathogens/Viral pathogens"
    ],
    [
      "plos",
      "/Medicine and health sciences/Pulmonology/Respiratory disorders/Respiratory infections"
    ],
    [
      "plos",
      "/Physical sciences/Physics/Condensed matter physics/Phase transitions/Melting"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/Artificial gene amplification and extension/Polymerase chain reaction"
    ]
  ],
  "openalex": "W7211992948",
  "pdf_url": "https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0357377&type=printable",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-09",
  "publisher": "Public Library of Science",
  "ris": "https://research.draconitic.com/paper/7fyfmfvckj1z6.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:56Z",
      "native_id": "10.1371/journal.pone.0357377",
      "source": "plos",
      "url": "https://doi.org/10.1371/journal.pone.0357377"
    }
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  "subjects": [
    {
      "field": "biology",
      "origin": "plos:microbiology",
      "subfield": "microbiology"
    }
  ],
  "title": "Development of a multiplex real-time PCR assay with fluorescence probe-melting-curve analysis for one-tube detection of respiratory pathogens",
  "title_text": "Development of a multiplex real-time PCR assay with fluorescence probe-melting-curve analysis for one-tube detection of respiratory pathogens",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:17:56Z",
  "url": "https://research.draconitic.com/paper/7fyfmfvckj1z6",
  "venue": "PLOS One",
  "versions": []
}