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Background <p>Picornaviruses, common infectious agents in humans and various animal species, pose significant health threats. Conventional monoplex PCR is widely employed in laboratory diagnostics but is relatively time-intensive and laborious.</p> Results <p>In this study, we developed a multiplex <i>Taq</i>Man probe-based real-time quantitative PCR (qPCR) assay for the rapid and simultaneous detection of kobuvirus, parechovirus B, rosavirus B and hunnivirus in murine rodent and shrew samples. The approach demonstrated high sensitivity and specificity, with detection limits of 1 × 10<sup>2</sup> copies/µL for kobuvirus, parechovirus B, and rosavirus B, and 50 copies/µL for hunnivirus. Evaluation using 149 clinical samples showed strong concordance with conventional PCR methods.</p> Conclusions <p>This work developed an effective multiplex qPCR method for the simultaneous detection of emerging picornaviruses particularly in rodents, including kobuvirus, parechovirus B, rosavirus B, and hunnivirus. Our findings contribute valuable insights into the monitoring and prevention of zoonotic diseases associated with these pathogens.</p>

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A TaqMan probe-based multiplex real-time quantitative pcr for simultaneous detection of kobuvirus, parechovirus B, rosavirus B, and hunnivirus carried by murine rodents and shrews

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年3月6日

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  • Shunchang Fan,
  • Minyi Zhang,
  • Yucheng Li,
  • Jingli Tian,
  • Juxian Xian,
  • Qing Chen

摘要

Background

Picornaviruses, common infectious agents in humans and various animal species, pose significant health threats. Conventional monoplex PCR is widely employed in laboratory diagnostics but is relatively time-intensive and laborious.

Results

In this study, we developed a multiplex TaqMan probe-based real-time quantitative PCR (qPCR) assay for the rapid and simultaneous detection of kobuvirus, parechovirus B, rosavirus B and hunnivirus in murine rodent and shrew samples. The approach demonstrated high sensitivity and specificity, with detection limits of 1 × 102 copies/µL for kobuvirus, parechovirus B, and rosavirus B, and 50 copies/µL for hunnivirus. Evaluation using 149 clinical samples showed strong concordance with conventional PCR methods.

Conclusions

This work developed an effective multiplex qPCR method for the simultaneous detection of emerging picornaviruses particularly in rodents, including kobuvirus, parechovirus B, rosavirus B, and hunnivirus. Our findings contribute valuable insights into the monitoring and prevention of zoonotic diseases associated with these pathogens.