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<p>Bovine viral diarrhea virus (BVDV), <i>Pasteurella multocida</i> (PM), and <i>Brucella spp.</i> are major pathogens that significantly compromise the global cattle industry. To address the need for accurate molecular detection, a novel triplex TaqMan qPCR assay was established targeting three conserved genetic markers: BVDV 5’UTR, PM <i>Kmt1</i>, and Brucella <i>Bcsp31</i>. The method achieved detection limits of 18.9, 3.19, and 3.32 copies/µL for BVDV, PM and <i>Brucella spp.</i>, with acceptable amplification efficiencies (93–101%). Additionally, the assay was highly specific, as no cross-reactions were detected with a panel of non-target bovine pathogens. Performance evaluation confirmed high reliability, with intra- and inter-assay CVs consistently under 1.34%. Large-scale epidemiological surveillance of cattle suspected of BVDV, PM, and <i>Brucella spp.</i> infections in Guizhou Province of China from 2022 to 2025 revealed overall detection rates of 18.47% for BVDV, 20.76% for PM, and 3.63% for <i>Brucella spp.</i> from nasal swabs. Co-infections were also observed, most notably BVDV and PM (3.37%), followed by PM and <i>Brucella spp.</i> (0.67%), and BVDV and <i>Brucella spp.</i> (0.31%). Annual detection rates across the four-year period remained relatively stable, with slight upward fluctuations for all three pathogens. This study developed an efficient triplex qPCR platform for the surveillance and differential detection of these high-impact cattle pathogens.</p>

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Epidemiological surveillance of BVDV, Pasteurella multocida, and Brucella spp. infections in cattle from Guizhou, China using a newly established triplex TaqMan qPCR assay

  • 期刊论文

  • 发表日期：

    2026年5月11日

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  • Jingjin Hu,
  • Weijie Zhou,
  • Shaobo Liang,
  • Xiaoyue Wang,
  • Feng Pang

摘要

Bovine viral diarrhea virus (BVDV), Pasteurella multocida (PM), and Brucella spp. are major pathogens that significantly compromise the global cattle industry. To address the need for accurate molecular detection, a novel triplex TaqMan qPCR assay was established targeting three conserved genetic markers: BVDV 5’UTR, PM Kmt1, and Brucella Bcsp31. The method achieved detection limits of 18.9, 3.19, and 3.32 copies/µL for BVDV, PM and Brucella spp., with acceptable amplification efficiencies (93–101%). Additionally, the assay was highly specific, as no cross-reactions were detected with a panel of non-target bovine pathogens. Performance evaluation confirmed high reliability, with intra- and inter-assay CVs consistently under 1.34%. Large-scale epidemiological surveillance of cattle suspected of BVDV, PM, and Brucella spp. infections in Guizhou Province of China from 2022 to 2025 revealed overall detection rates of 18.47% for BVDV, 20.76% for PM, and 3.63% for Brucella spp. from nasal swabs. Co-infections were also observed, most notably BVDV and PM (3.37%), followed by PM and Brucella spp. (0.67%), and BVDV and Brucella spp. (0.31%). Annual detection rates across the four-year period remained relatively stable, with slight upward fluctuations for all three pathogens. This study developed an efficient triplex qPCR platform for the surveillance and differential detection of these high-impact cattle pathogens.