Development of an Effective Double Antigen Sandwich ELISA Based on p30 Protein to Detect Antibodies against African Swine Fever Virus

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 tar...

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Bibliographic Details
Published in:Viruses 2022-09, Vol.14 (10), p.2170
Main Authors: Wang, Mengxiang, Song, Jinxing, Sun, Junru, Du, Yongkun, Qin, Xiaodong, Xia, Lu, Wu, Yanan, Zhang, Gaiping
Format: Article
Language:English
Subjects:
African swine fever
African swine fever virus
Antibodies
Antigens
Asfarviridae
diagnosis
DNA viruses
double-antigen sandwich ELISA
E coli
Enzyme-linked immunosorbent assay
Enzymes
Foot & mouth disease
Health aspects
Hogs
Horseradish peroxidase
Immunogenicity
Infectious diseases
Methods
p30 Protein
Physiological aspects
prokaryotic expression system
Proteins
Reproducibility
Serology
Statistical analysis
Viral antibodies
Viral infections
Viral proteins
Viruses
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Summary: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.
ISSN:1999-4915
1999-4915
DOI:10.3390/v14102170