A Detection Method of African Swine Fever Virus based on Enzymatic Recombinase Amplification
This experiment was conducted to establish a rapid nucleic acid detection method for African swine fever virus(ASFV)based on enzymatic recombinase amplification(ERA).We designed ERA probes and primers specific for the conserved sequence of ASFV B646L gene,and optimized the reaction conditions in order to establish an ERA method for the detection of ASFV DNA under isothermal conditions.The ERA method for ASFV demonstrated high specificity and no cross-reaction with other pathogens;the CV was less than 10%,indicative of good reproducibility.The lowest detection limit for the ERA method is 85 copies·μL-1;comparison with the World Organization for Animal Health(WOAH)recommended qPCR diagnostic method for African swine fever(ASF)demonstrated a Kappa value of 0.961,suggestive of high identity with African swine fever qPCR diagnostic method.ERA-based method for the detection of ASFV can be used for the rapid detection of ASFV in the field.