Rapid detection of ASFV, PDCoV, and SVA in pork and its products by recombinant enzyme polymerase amplification

ObjectiveTo establish a rapid detection method for multiple recombinant enzyme polymerase amplification （RPA） of African swine fever virus （ASFV）， Porcine Delta coronavirus （PDCoV）， and Seneca virus type A （SVA）.MethodsRPA detection was carried out using recombinant plasmid as a template， and reaction conditions such as temperature， time， and primer probe ratio were optimized. Multiple RPA detection methods were established， which were applied to the detection of actual samples， and the accuracy of the established method was evaluated by comparing with the results of real-time fluorescence polymerase chain reaction （qPCR）.ResultsSimultaneous detection of the three viruses can be achieved within 20 min with good specificity. Sensitivity to ASFV， PDCoV， and SVA was 940， 770， and 570 copies/μL， respectively. The accuracy rates for actual sample detection were 93.33%， 100.00%， and 100.00%， respectively.ConclusionThe established multiple RPA method is fast and convenient， and it is suitable for on-site preliminary