Chen Wenxian, Niu Xinni, Zeng Weijun, Fang Yiqi, Zhu Zixiang, Yi Lin, Zhao Mingqiu, Ding Hongxing, Fan Shuangqi, Li Zhaoyao, Chen Jinding
College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou Veterinary Research Institute, Lanzhou University, Chinese Academy of Agricultural Sciences, Lanzhou, China.
Front Vet Sci. 2025 Jun 23;12:1526005. doi: 10.3389/fvets.2025.1526005. eCollection 2025.
Foot-and-mouth disease virus (FMDV) causes blister-like lesions in animals, leading to significant economic losses in animal husbandry. Accurate and rapid detection of FMD is crucial for effective prevention and control. The recombinase-aided amplification (RAA) technique enables rapid amplification of target fragments under isothermal conditions. In this study, based on the conserved sequence of the FMDV 3D gene, optimal RAA primers and probes were designed and screened, and a simple, rapid, and visual FMDV nucleic acid RAA test strip was developed. The optimum reaction conditions of the assay were determined to be 32°C for 30 min. The specificity, sensitivity, and repeatability of the FMDV nucleic acid RAA test strip were evaluated. 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. The lowest detection limits for recombinant plasmids and virus titer were 10 copies/μL and 10 TCID/mL, respectively. In addition, all 17 positive samples and 21 negative samples were accurately identified using the FMDV nucleic acid RAA test strip, resulting in a 100% positive detection rate. In conclusion, the FMDV nucleic acid RAA test strip developed in this study-characterized by high specificity, efficiency, and sensitivity-offers a robust technical platform for the prevention and control of FMD.
口蹄疫病毒（FMDV）可导致动物出现水泡样病变，给畜牧业造成重大经济损失。准确快速地检测口蹄疫对于有效防控至关重要。重组酶辅助扩增（RAA）技术可在等温条件下快速扩增靶片段。本研究基于口蹄疫病毒3D基因的保守序列，设计并筛选了最佳RAA引物和探针，研制出一种简单、快速、可视化的口蹄疫病毒核酸RAA检测试纸条。确定该检测方法的最佳反应条件为32℃反应30分钟。对口蹄疫病毒核酸RAA检测试纸条的特异性、灵敏度和重复性进行了评估。结果表明，口蹄疫病毒核酸RAA检测试纸条与口蹄疫病毒核酸特异性反应，与其他病毒无交叉反应。重组质粒和病毒滴度的最低检测限分别为10拷贝/μL和10 TCID/mL。此外，使用口蹄疫病毒核酸RAA检测试纸条对17份阳性样品和21份阴性样品全部准确鉴定，阳性检出率达100%。综上所述，本研究研制的口蹄疫病毒核酸RAA检测试纸条具有高特异性、高效性和灵敏度，为口蹄疫的防控提供了一个强大的技术平台。
