We established a recombinase polymerase amplification (RPA) assay for the rapid detection of viral hemorrhagic septicemia virus (VHSV) using primers designed to the N gene in this species. Optimization experiments showed that the optimal amplification temperature of the RPA assay was 37 °C and that the reaction could be completed within only 15 min. Specificity tests showed that the RPA assay did not exhibit any cross-reactivity with other infective fish viruses. Sensitivity tests further showed that the detection limit of the new RPA assay was 8.3 copies/μL, indicating that this assay was more sensitive than traditional RT-PCR method. The method was verified by detecting VHSV in 1924 batches of samples collected from domestic and imported fishes. The detection results were consistent with that of traditional RT-PCR, and the specificity and sensitivity of the method met the detection requirements for aquatic animal diseases. Collectively, our findings indicate that the novel RPA assay is fast, simple, specific, sensitive, and has significant potential for the clinic, on-site testing, and a wide range of other applications.

我们建立了重组酶聚合酶扩增（RPA）测定法，以使用针对该物种N基因的引物快速检测病毒性出血性败血病病毒（VHSV）。优化实验表明，RPA分析的最佳扩增温度为37°C，反应仅需15分钟即可完成。特异性测试表明，RPA分析与其他感染性鱼病毒没有任何交叉反应。敏感性测试进一步表明，新的RPA测定的检测限为8.3拷贝/μL，表明该测定比传统的RT-PCR方法更灵敏。该方法通过检测1924批次的国产和进口鱼类样品中的VHSV进行了验证。检测结果与传统RT-PCR一致，该方法的特异性和敏感性满足水生动物疾病的检测要求。总的来说，我们的发现表明，新颖的RPA测定法快速，简单，特异，灵敏，在临床，现场测试以及其他广泛应用中具有巨大潜力。