African swine fever has caused huge losses to the global pig industry. In the absence of effective vaccines, reliable detection methods are crucial. The p30 protein of ASFV is often used as a target for detection due to its high antigenicity in the early stage of virus replication. In this study, we modified the p30 protein using magnetic nanoparticles to increase its specific surface area and suspension in solution. Based on this modified p30 protein, we optimized the experimental steps to determine the best reaction conditions and then established a double probe competitive ELISA method. The established detection method can produce results within 45 min, with repeatability between and within batches <15 %, and it does not react with other pathogens, which demonstrates high specificity. In conclusion, we modified the ASFV p30 protein with magnetic nanoparticles and established a rapid, ultra-sensitive, and convenient detection method for ASF based on the modified p30. This also provides a new reference for the rapid detection of other veterinary diseases.
Keywords: African swine fever; Double probe detection method; Magnetic nanoparticles; Rapid diagnosis; p30 protein.
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