A novel rapid detection of Senecavirus A using recombinase polymerase amplification (RPA) coupled with lateral flow (LF) dipstrip.

SENECAVIRUS A: (SVA), an emerging picornavirus, has been associated with vesicular disease and neonatal mortality in swine, posing a great threat to the global swine industry. Accurate diagnosis of SVA is crucial for the effective prevention and control disease. In the present study, a simple, rapid and accurate diagnostic assay was developed combining recombinase polymerase amplification and a lateral flow dipstrip (RPA-LF) to detect SVA infection. Using recombinant plasmid pMD19-T-VP1 DNA as a template, the RPA-LF optimal reaction conditions were incubated at 35 °C for 25 min, and the result was visualized directly on the dipstrip. The specificity assay showed no cross-reactivity with other tested viruses, and the sensitivity assay revealed the minimum detection limit was 15 copies/μl. Moreover, the RPA-LF method was successfully applied with viral cDNA as template to test clinical samples, with no significant difference being observed between RPA-LF and qRT-PCR. Hence, the established RPA-LF assay could be used as a potential optional rapid, reliable, sensitive and low-cost method for field diagnosis of SVA, especially in resource-limited regions.