Viruses, Vol. 15, Pages 1939: Developing an Indirect ELISA for the Detection of African Swine Fever Virus Antibodies Using a Tag-Free p15 Protein Antigen
Viruses, Vol. 15, Pages 1939: Developing an Indirect ELISA for the Detection of African Swine Fever Virus Antibodies Using a Tag-Free p15 Protein Antigen
Viruses doi: 10.3390/v15091939
Authors:
Zhi Wu
Huipeng Lu
Dewei Zhu
Jun Xie
Fan Sun
Yan Xu
Hua Zhang
Zhijun Wu
Wenlong Xia
Shanyuan Zhu
African swine fever (ASF) is one of the most severe diseases caused by the ASF virus (ASFV), causing massive economic losses to the global pig industry. Serological tests are important in ASF epidemiological surveillance, and more antigen targets are needed to meet market demand for ASFV antibody detection. In the present study, ASFV p15 protein was fusion-expressed in Escherichia coli (E. coli) with elastin-like polypeptide (ELP), and the ELP-p15 protein was purified using a simple inverse transition cycling (ITC) process. The ELP tag was cleaved off using tobacco etch virus protease (TEVp), resulting in a tag-free p15 protein. Western blot analysis demonstrated that the p15 protein reacted strongly with ASFV-positive serum. The p15 protein was used as a coating antigen in an indirect ELISA (iELISA) for detecting ASFV antibodies. The p15-iELISA method demonstrated high specificity to ASFV-positive sera, with a maximum detection dilution of 1:1600. Moreover, the method exhibited good reproducibility, with less intra-assay and inter-assay CV values than 10%. Therefore, p15-iELISA offers a novel approach for accurately detecting ASFV antibodies with significant clinic...
Source: Viruses - Category: Virology Authors: Zhi Wu Huipeng Lu Dewei Zhu Jun Xie Fan Sun Yan Xu Hua Zhang Zhijun Wu Wenlong Xia Shanyuan Zhu Tags: Article Source Type: research