Development of a novel double-antibody sandwich quantitative ELISA for detecting SADS-CoV infection
文献类型： 外文期刊
第一作者： Cao, Liyan
作者： Cao, Liyan;Kong, Xiangyu;Zhang, Yu;Suo, Xuepeng;Li, Xiangtong;Duan, Yueyue;Yuan, Cong;Wang, Qi;Cao, Liyan;Kong, Xiangyu;Zhang, Yu;Suo, Xuepeng;Li, Xiangtong;Duan, Yueyue;Yuan, Cong;Zheng, Haixue;Wang, Qi;Cao, Liyan;Kong, Xiangyu;Zhang, Yu;Suo, Xuepeng;Li, Xiangtong;Duan, Yueyue;Yuan, Cong;Wang, Qi
作者机构：
关键词： Swine acute diarrhea syndrome coronavirus (SADS-CoV); Nucleocapsid protein; Monoclonal antibody; DAS-qELISA; Antigen detection
期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：5.0； 五年影响因子：5.2 ）
ISSN： 0175-7598
年卷期： 2023 年 107 卷 7-8 期
页码：
收录情况： SCI
摘要： Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an emerging swine enteric alphacoronavirus that can cause acute diarrhea, vomiting, dehydration, and death of newborn piglets. In this study, we developed a double-antibody sandwich quantitative enzyme-linked immunosorbent assay (DAS-qELISA) for detection of SADS-CoV by using an anti-SADS-CoV N protein rabbit polyclonal antibody (PAb) and a specific monoclonal antibody (MAb) 6E8 against the SADS-CoV N protein. The PAb was used as the capture antibodies and HRP-labeled 6E8 as the detector antibody. The detection limit of the developed DAS-qELISA assay was 1 ng/mL of purified antigen and 10(1.08)TCID(50)/mL of SADS-CoV, respectively. Specificity assays showed that the developed DAS-qELISA has no cross-reactivity with other swine enteric coronaviruses, such as porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and porcine deltacoronavirus (PDCoV). Three-day-old piglets were challenged with SADS-CoV and collected anal swab samples which were screened for the presence of SADS-CoV by using DAS-qELISA and reverse transcriptase PCR (RT-PCR). The coincidence rate of the DAS-qELISA and RT-PCR was 93.93%, and the kappa value was 0.85, indicating that DAS-qELISA is a reliable method for applying antigen detection of clinical samples.
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