Abstract
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.
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