ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Veterinary Infectious Diseases
Volume 12 - 2025 | doi: 10.3389/fvets.2025.1601685
Responses to Editor (Manuscript ID: 1601685 )
Provisionally accepted
- Zhejiang Academy of Agricultural Sciences, Hangzhou, China
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Avipoxvirus (APV), a DNA virus, is widely prevalent in avian species, causing clinical symptoms of fowlpox and resulting in reduced egg production, slower broiler growth, and increased mortality. Its spread poses a serious threat to the global poultry industry, potentially leading to significant economic losses. To effectively control the spread of APV, particularly its major species, fowlpoxvirus and pigeonpoxvirus, developing rapid and specific diagnostic tools is critical. In this study, a novel multi-enzyme isothermal rapid amplification (MIRA) assay was developed and evaluated for sensitivity, specificity and applicability. By screening various primer-probe combinations, an optimal pair was identified targeting a conserved region of the viral P4b gene for APV detection. The MIRA assay operates at a constant temperature and provides results within 15 minutes, then results can be visualized through fluorescence signal detection or observed under blue light illumination. It demonstrated high specificity with no cross-reactivity with other avian pathogens and achieved a detection limit of 50 copies/μl, consistent with the quantitative PCR (qPCR) assay. Further evaluation with 86 clinical samples showed that the accuracy of the MIRA was comparable to that of qPCR in detecting fowlpoxvirus and pigeonpoxvirus. These results highlight the convenience, sensitivity, and rapidity of the MIRA assay as a promising tool for diagnosing APV.
Keywords: Avipoxvirus, multienzyme isothermal amplification, Nucleic acid detection, Rapid assay, clinic diagnostic
Received: 28 Mar 2025; Accepted: 18 Apr 2025.
Copyright: © 2025 Zhu, Huo, Chen, Yuan, Hua, Yun, Zhang, Ni and Ye. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Yinchu Zhu, Zhejiang Academy of Agricultural Sciences, Hangzhou, China
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