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Development of an Effective Double Antigen Sandwich ELISA Based on p30 Protein to Detect Antibodies against African Swine Fever Virus
Authors:Mengxiang Wang  Jinxing Song  Junru Sun  Yongkun Du  Xiaodong Qin  Lu Xia  Yanan Wu  Gaiping Zhang
Affiliation:1.College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China;2.International Joint Research Center of National Animal Immunology, Zhengzhou 450046, China;3.Longhu Laboratory, Zhengzhou 450046, China;4.Henan Engineering Laboratory of Animal Biological Products, Zhengzhou 450046, China
Abstract:African swine fever (ASF), the highly lethal swine infectious disease caused by the African swine fever virus (ASFV), is a great threat to the swine industry. There is no effective vaccine or diagnostic method to prevent and control this disease currently. The p30 protein of ASFV is an important target for serological diagnosis, expressed in the early stage of viral replication and has high immunogenicity and sequence conservatism. Here, the CP204L gene was cloned into the expression vector pET-30a (+), and the soluble p30 protein was successfully expressed in the E. coli prokaryotic expression system and then labeled with horseradish peroxidase (HRP) to be the enzyme-labeled antigen. Using the purified recombinant p30 protein, a double-antigen sandwich ELISA for ASFV antibody detection was developed. This method exhibits excellent specificity, sensitivity and reproducibility in clinical sample detection with lower cost and shorter production cycles. Taken together, this study provides technical support for antibody detection for ASFV.
Keywords:African swine fever virus   double-antigen sandwich ELISA   diagnosis   p30 protein   prokaryotic expression system
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