{
  "abstract": "Bovine respiratory disease (BRD) is a clinicopathological syndrome caused by complex interactions among host and environmental factors, viruses and bacteria. The multi-agent BRD etiology makes laboratory diagnosis challenging, as various pathogens should be investigated for an accurate results. Herein, we describe an endpoint multiplex PCR/RT-PCR for detection of major BRD-associated viruses: bovine herpesvirus 1 (BoAHV-1), bovine pestiviruses (BVDV-1, BVDV-2, HoBiPeV), bovine parainfluenza virus 3 (BPIV-3), bovine respiratory syncytial virus (BRSV) and bovine coronavirus (BCoV). We designed high-coverage and target-specific primers to be properly combined in the multiplex assay. Following standardization of individual reactions, the multiplex PCR/RT-PCR was optimized for reaction conditions and mix composition. The amplicons of the five targets were easily differentiated in agarose gel: 121 bp (BPIV-3), 157 bp (BoAHV-1), 204 bp (pestiviruses), 282 bp (BRSV) and 323 bp (BCoV). The limits of detection (LoDs) of our multiplex were 38 TCID₅₀ (BoAHV-1 and BPIV-3), 16 TCID₅₀ (pestiviruses), 13 TCID₅₀ (BRSV) and a 10- 2 dilution of a pool of five BCoV positive samples. No amplification of the main bacteria (Pasteurella multocida; Mannheimia haemolytica and Histophilus somni) potentially involved in BRD was observed. Finally, 289 respiratory specimens from animals with respiratory/nasal signs were evaluated by individual and multiplex PCR/RT-PCR tests to assess the diagnostic performance of the assay. Both tests resulted in 14.8% positive samples (43/289) being 19 for BPIV-3 (19/43), ten for BCoV (10/43), seven for BRSV (7/43), six for BoAHV-1 (6/43) and one for pestiviruses (1/43). Overall, our multiplex PCR/RT-PCR assay represents an alternative for an easier, faster and more practical laboratory diagnosis of viruses associated with BRD.",
  "abstract_shown": true,
  "affiliations": [
    "Setor de Virologia, Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, Av. Roraima, 1000, Prédio 63 A, Camobi, Santa Maria, Rio Grande do Sul, 97105-900, Brazil. nataliahpedroso@gmail.com.",
    "Programa de Pós-graduação em Medicina Veterinária, Universidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul, Brazil. nataliahpedroso@gmail.com.",
    "Setor de Virologia, Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, Av. Roraima, 1000, Prédio 63 A, Camobi, Santa Maria, Rio Grande do Sul, 97105-900, Brazil.",
    "Departamento de Microbiologia e Parasitologia, Universidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul, Brazil.",
    "Laboratório NB3 de Neuroimunologia, Universidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul, Brazil.",
    "Programa de Pós-graduação em Farmacologia, Universidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul, Brazil.",
    "Setor de Virologia, Instituto Keizo Asami, Universidade Federal de Pernambuco, Pernambuco, Brazil.",
    "Programa de Pós-graduação em Medicina Veterinária, Universidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul, Brazil.",
    "Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, Brasil."
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 7,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Natália Hettwer Pedroso Alessio",
      "orcid": "0000-0002-7826-0457"
    },
    {
      "affs": [
        2,
        3,
        4,
        5,
        6
      ],
      "name": "José Valter Joaquim Silva Júnior"
    },
    {
      "affs": [
        2,
        7
      ],
      "name": "Iasmim Flôres Dionisio"
    },
    {
      "affs": [
        2
      ],
      "name": "Vinicius Soares Dos Santos"
    },
    {
      "affs": [
        8
      ],
      "name": "Viviani Gomes"
    },
    {
      "affs": [
        2
      ],
      "name": "Rudi Weiblen"
    },
    {
      "affs": [
        2
      ],
      "name": "Eduardo Furtado Flores"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/awt9bzwn58hf2.bib",
  "cited_by": 0,
  "doi": "10.1007/s42770-026-02097-2",
  "fulltext_url": "https://europepmc.org/articles/PMC13728016",
  "id": "awt9bzwn58hf2",
  "issns": [
    "1517-8382",
    "1678-4405"
  ],
  "landing_url": "https://europepmc.org/article/MED/42970197",
  "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": [
    [
      "heading",
      "Veterinary and Animal Production - Research Paper"
    ],
    [
      "openalex",
      "1103|Animal Virus Infections Studies"
    ]
  ],
  "openalex": "W7225080409",
  "pdf_url": "https://europepmc.org/articles/PMC13728016?pdf=render",
  "pmcid": "PMC13728016",
  "pmid": "42970197",
  "published": "2026-11-02",
  "publisher": "Brazilian Society of Microbiology",
  "ris": "https://research.draconitic.com/paper/awt9bzwn58hf2.ris",
  "sources": [
    {
      "first_seen": "2026-11-04T15:20:43Z",
      "native_id": "MED:42970197",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42970197"
    },
    {
      "first_seen": "2026-11-06T12:23:47Z",
      "native_id": "PMC13728016",
      "source": "pmc",
      "url": "https://doi.org/10.1007/s42770-026-02097-2"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "openalex:1103|Animal Virus Infections Studies",
      "subfield": "zoology-behaviour"
    }
  ],
  "title": "An endpoint multiplex PCR/RT-PCR for detection of viral agents of bovine respiratory disease.",
  "title_text": "An endpoint multiplex PCR/RT-PCR for detection of viral agents of bovine respiratory disease.",
  "type": "peer-reviewed",
  "updated_at": "2026-11-07T12:21:05Z",
  "url": "https://research.draconitic.com/paper/awt9bzwn58hf2",
  "venue": "Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]",
  "versions": []
}