Development of ERA detection method for African swine fever virus based on B646L, EP402R and MGF360/505 genes

Objective  To establish a rapid detection method of African swine fever virus (ASFV) DNA based on enzymatic recombinase amplification (ERA) technology.

Method  Specific ERA probes and primers were designed according to the conserved sequences of B646L and EP402R genes, and MGF360/505 gene which was one of the ASFV multigene family members. Through optimizing reaction conditions, the ERA method for detecting ASFV DNA was finally established under the isothermal condition of 42 ℃.

Result  The detection results of three methods of ERA-ASFV-B646L, ERA-ASFV-EP402R and ERA-ASFV-MGF could be obtained within 16, 7 and 13 min respectively. The copy number detection limitation of positive sample was all 10² μL⁻¹. The coincidence rate was 100% compared with the common method in the technical standard of ASFV diagnosis in China.

Conclusion  The established ERA method can be used for rapid detection of ASFV, and provides an effective rapid detection method for epidemiological investigation and field detection.