A rapid and visual detection assay for Senecavirus A based on recombinase-aided amplification and lateral flow dipstick
CORRECTION article
A rapid and visual detection assay for Senecavirus A based on recombinase-aided amplification and lateral flow dipstick
Provisionally accepted- 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 Group Academy, Wen’s Foodstuffs Group Co., Ltd., Guangzhou, China
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 3.86×101 copies/µL, 8.76×10-7 ng/µL, and 1×100.25 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 suited for utilization in lack-instrumented laboratory and field settings.
Keywords: SenecavirusA, Recombinase-aided amplification, Lateral flow dipstick, Sensitivity, specificity, visual detection
Received: 12 Nov 2024; Accepted: 15 Nov 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 Group Academy, 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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