Wang Yan, Shang Jinyuan, Li Zhijie, Zhang Ao, Cheng Yuening
Key Laboratory of Economic Animal Diseases, Ministry of Agriculture, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, China.
Front Vet Sci. 2024 Mar 27;11:1360504. doi: 10.3389/fvets.2024.1360504. eCollection 2024.
Bovine Viral Diarrhea Virus (BVDV) and Infectious Bovine Rhinotracheitis Virus (IBRV) are the two most prevalent infectious diseases in cattle. They both can cause persistent infection and immunosuppression, resulting in significant economic losses in the livestock industry. Therefore, rapid detection of early BVDV and IBRV infections is crucial. In this study, a method for the rapid detection of BVDV and IBRV was established by using recombinase polymerase amplification (RPA) combined with lateral flow device (LFD). By optimizing the temperature and time conditions of the RPA reaction, the sensitivity, specificity, and clinical performance were evaluated. The results indicated that the RPA reaction could be completed at 40°C within 25 min. The LOD for BVDV and IBRV by RPA-LFD were 5.1 × 10 copies/μL and 6.65 × 10 copies/μL, respectively, with no cross-reactivity observed with other viruses such as CSFV, BRSV, BPIV3, BRV, and BCoV. Testing of 32 clinical samples showed consistent results between RPA-LFD and qPCR. The RPA-LFD method established in this study can be used for the rapid clinical detection of BVDV and IBRV, which providing a rapid and convenient molecular biology approach for on-site rapid detection and epidemiological investigations. Simultaneously, it offers technical support for the prevention and control of these viruses.
牛病毒性腹泻病毒（BVDV）和牛传染性鼻气管炎病毒（IBRV）是牛群中两种最普遍的传染病。它们都可引起持续性感染和免疫抑制，给畜牧业造成重大经济损失。因此，快速检测早期BVDV和IBRV感染至关重要。在本研究中，通过重组酶聚合酶扩增（RPA）结合侧向流动装置（LFD）建立了一种快速检测BVDV和IBRV的方法。通过优化RPA反应的温度和时间条件，评估了其敏感性、特异性和临床性能。结果表明，RPA反应可在40°C下25分钟内完成。RPA-LFD对BVDV和IBRV的检测限分别为5.1×10拷贝/μL和6.65×10拷贝/μL，与其他病毒如猪瘟病毒（CSFV）、牛呼吸道合胞病毒（BRSV）、牛副流感病毒3型（BPIV3）、牛轮状病毒（BRV）和牛冠状病毒（BCoV）未观察到交叉反应。对32份临床样本的检测显示，RPA-LFD和qPCR结果一致。本研究建立的RPA-LFD方法可用于BVDV和IBRV的快速临床检测，为现场快速检测和流行病学调查提供了一种快速便捷的分子生物学方法。同时，为这些病毒的防控提供了技术支持。
