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ORIGINAL RESEARCH article

Front. Cell. Infect. Microbiol.
Sec. Veterinary and Zoonotic Infection
Volume 14 - 2024 | doi: 10.3389/fcimb.2024.1474676
This article is part of the Research Topic Viral Diseases in Swine View all articles

A rapid and visual detection assay for Senecavirus A based on recombinase-aided amplification and lateral flow dipstick

Provisionally accepted
Yiwan Song Yiwan Song 1,2Yiqi Fang Yiqi Fang 1,2*Shuaiqi Zhu Shuaiqi Zhu 1,2*Weijun Wang Weijun Wang 1*Lianxiang Wang Lianxiang Wang 3*Wenxian Chen Wenxian Chen 1,2*Yintao He Yintao He 1,2*Lin Yi Lin Yi 1,2*Hongxing Ding Hongxing Ding 1,2*Mingqiu Zhao Mingqiu Zhao 1*Shuangqi Fan Shuangqi Fan 1,2*Zhaoyao Li Zhaoyao Li 1,3*Jinding Chen Jinding Chen 1,2*
  • 1 College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
  • 2 Key Laboratory of Prevention and Control of Zoonotic Diseases of Guangdong Province, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
  • 3 Wen’s Foodstuffs Group Co., Ltd, Guangzhou, China

The final, formatted version of the article will be published soon.

    Background: Senecavirus A (SVA) is a newly pathogenic virus correlated with the acute death of piglets and vesicular lesions in pigs. The further prevalence of SVA will cause considerable economic damage to the global pig farming industry. Therefore, rapid and accurate diagnostic tools for SVA are crucial for preventing and controlling the disease.We designed multiple primer pairs targeting the most conserved region of the SVA 3D gene and selected those with the highest specificity. Nfo-probes were subsequently developed based on the highest specificity primer pairs. Subsequently, the recombinase-assisted amplification (RAA) reaction was completed, and the reaction temperature and duration were optimized. The RAA amplicons were detected using a lateral flow device (LFD). Finally, a rapid and intuitive RAA-LFD assay was established against SVA.The SVA RAA-LFD assay can be performed under reaction conditions of 35°C within 17 minutes, with results observable to the naked eye. We then evaluated the performance of this method. It exhibited high specificity and no cross-reaction with the other common swine pathogens. The lowest detectable limits of this method for the plasmid of pMD18-SVA-3D, DNA amplification product, and viral were 3.86×10 1 copies/µL, 8.76×10 -7 ng/µL, and 1×10 0.25 TCID50/mL, respectively. A total of 44 clinical samples were then tested using the RAA-LFD, PCR, and RT-qPCR methods. The results demonstrated a consistent detection rate between the RAA-LFD and RT-qPCR assays.The SVA RAA-LFD assay developed in our study exhibits excellent specificity, sensitivity, and time-saving attributes, making it ideally suited for utilization in lack-instrumented laboratory and field settings.

    Keywords: Senecavirus A, Recombinase-aided amplification, Lateral flow dipstick, Sensitivity, specificity, visual detection

    Received: 02 Aug 2024; Accepted: 01 Oct 2024.

    Copyright: © 2024 Song, Fang, Zhu, Wang, Wang, Chen, He, Yi, Ding, Zhao, 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:
    Yiqi Fang, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Shuaiqi Zhu, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Weijun Wang, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Lianxiang Wang, Wen’s Foodstuffs Group Co., Ltd, Guangzhou, China
    Wenxian Chen, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Yintao He, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Lin Yi, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Hongxing Ding, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Mingqiu Zhao, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Shuangqi Fan, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Zhaoyao Li, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
    Jinding Chen, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China

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