ORIGINAL RESEARCH article

Front. Vet. Sci.

Sec. Veterinary Epidemiology and Economics

Volume 12 - 2025 | doi: 10.3389/fvets.2025.1526005

Development of Nucleic Acid RAA Test Strips Assay for Rapid Detection of Foot-and-mouth Disease Virus

Provisionally accepted
Wenxian ChenWenxian Chen1Niu XinniNiu Xinni1Weijun ZengWeijun Zeng1Yiqi FangYiqi Fang1Zixiang ZhuZixiang Zhu2Lin YiLin Yi1Mingqiu ZhaoMingqiu Zhao1Hongxing DingHongxing Ding1Shuangqi FanShuangqi Fan1Zhaoyao LiZhaoyao Li3*Jinding ChenJinding Chen1*
  • 1College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, China
  • 2State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, China
  • 3Wen's Foodstuffs Group Co., Ltd., Xinxing, Yunfu, China

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FMDV causes blister-like lesions in animals, leading to significant economic losses in animal husbandry. It is of great significance for FMD prevention and control to detect FMDV accurately and rapidly. Recombinase aided amplification (RAA) technique enables rapid amplification of target fragments under isothermal conditions. In this paper, according to the conserved sequence of FMDV 3D gene, the best RAA primers and probes were designed and screened, and a simple and rapid visual FMDV nucleic acid RAA test strip was developed. It was proved that the optimum reaction condition of the method was 32℃ for 30 min. The specificity, sensitivity and repeatability of FMDV nucleic acid RAA test strip was analyzed in turn. The results demonstrated that the FMDV nucleic acid RAA test strip specifically reacted with FMDV nucleic acid and exhibited no cross-reactivity with other viruses, and the lowest detection limit of recombinant plasmids and virus titer was 10 copies/μL and 10 0 TCID50/mL, respectively. In addition, all 17 positive samples and 21 negative samples were tested correctly using FMDV nucleic acid RAA test strip, which of the positive detection rate of this method was 100%. In conclusion, the FMDV nucleic acid RAA test strip developed in this study, characterized by high specificity, efficiency, and sensitivity, provides a robust technical platform for the prevention and control of FMD.

Keywords: Foot-and-Mouth Disease Virus, Isothermal amplification, Recombinase aided amplification, Nucleic acid test strip, Rapid detection

Received: 12 Nov 2024; Accepted: 04 Jun 2025.

Copyright: © 2025 Chen, Xinni, Zeng, Fang, Zhu, Yi, Zhao, Ding, Fan, Li and Chen. 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:
Zhaoyao Li, Wen's Foodstuffs Group Co., Ltd., Xinxing, Yunfu, China
Jinding Chen, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, Guangdong Province, China

Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Supplementary Material