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Currently, there are no suitable serological methods to assess the infection of SADS\u2010CoV and effectiveness of vaccines, making an urgent need to exploit effective enzyme\u2010linked immunosorbent assay (ELISA) to compensate for this deficiency. Here, a recombinant plasmid that expresses the spike (S) protein of SADS\u2010CoV fused to the Fc domain of human IgG was constructed to generate recombinant <jats:italic>baculovirus<\/jats:italic> and expressed in HEK 293F cells. The S\u2010Fc protein was purified with protein G Resin, which retained reactivity with anti\u2010human Fc and anti\u2010SADS\u2010CoV antibodies. The S\u2010Fc protein was then used to develop an indirect ELISA (S\u2010iELISA) and the reaction conditions of S\u2010iELISA were optimized. As a result, the cut\u2010off value was determined as 0.3711 by analyzing OD<jats:sub>450nm<\/jats:sub> values of 40 SADS\u2010CoV\u2010negative sera confirmed by immunofluorescence assay (IFA) and western blot. The coefficient of variation (CV) of 6 SADS\u2010CoV\u2010positive sera within and between runs of S\u2010iELISA were both less than 10%. The cross\u2010reactivity assays demonstrated that S\u2010iELISA was non\u2010cross\u2010reactive with other swine viruses' sera. Furthermore, the overall coincidence rate between IFA and S\u2010iELISA was 97.3% based on testing 111 clinical serum samples. Virus neutralization test with seven different OD<jats:sub>450nm<\/jats:sub> values of the sera showed that the OD<jats:sub>450nm<\/jats:sub> values tested by S\u2010iELISA are positively correlated with the virus neutralization assay. Finally, a total of 300 pig field serum samples were tested by S\u2010iELISA and commercial kits of other swine enteroviruses showed that the IgG\u2010positive for SADS\u2010CoV, TGEV, PDCoV and PEDV was 81.7, 54, 65.3 and 6%, respectively. The results suggest that this S\u2010iELISA is specific, sensitive, repeatable and can be applied for the detection of the SADS\u2010CoV infection in the swine industry.<\/jats:p>","DOI":"10.1111\/tbed.14196","type":"journal-article","created":{"date-parts":[[2021,6,20]],"date-time":"2021-06-20T05:19:22Z","timestamp":1624166362000},"page":"2065-2075","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Development of an indirect ELISA for detecting swine acute diarrhoea syndrome coronavirus IgG antibodies based on a recombinant spike protein"],"prefix":"10.1155","volume":"69","author":[{"given":"Peng","family":"Peng","sequence":"first","affiliation":[{"name":"State Key Laboratory of Biocontrol School of Life Science Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuepeng","family":"Gao","sequence":"additional","affiliation":[{"name":"Agricultural product Quality and Safety Inspection and Testing Center  Shenzhen China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingfeng","family":"Zhou","sequence":"additional","affiliation":[{"name":"Wen' s Group Academy Wen' s Foodstuffs Group Co, Ltd  Xinxing Guangdong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tianhua","family":"Jiang","sequence":"additional","affiliation":[{"name":"Wen' s Group Academy Wen' s Foodstuffs Group Co, Ltd  Xinxing Guangdong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shumei","family":"Zheng","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocontrol School of Life Science Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meiyan","family":"Huang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocontrol School of Life Science Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunyi","family":"Xue","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocontrol School of Life Science Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0311-5474","authenticated-orcid":false,"given":"Yongchang","family":"Cao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocontrol School of Life Science Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5697-5734","authenticated-orcid":false,"given":"Zhichao","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocontrol School of Life Science Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,6,29]]},"reference":[{"key":"e_1_2_10_2_1","first-page":"257","article-title":"A transmissible gastroenteritis in pigs","volume":"108","author":"Doyle L. 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