Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen

文献类型： 外文期刊

第一作者： Fan, Baochao

作者： Fan, Baochao;Sun, Jie;Zhu, Lin;Zhou, Jinzhu;Zhao, Yongxiang;Yu, Zhengyu;Sun, Bing;Guo, Rongli;He, Kongwang;Li, Bin;Fan, Baochao;Sun, Jie;Zhu, Lin;Zhou, Jinzhu;Zhao, Yongxiang;Yu, Zhengyu;Sun, Bing;Guo, Rongli;He, Kongwang;Li, Bin;Fan, Baochao;Sun, Jie;Zhu, Lin;Zhou, Jinzhu;Zhao, Yongxiang;Yu, Zhengyu;Sun, Bing;Guo, Rongli;He, Kongwang;Li, Bin;Fan, Baochao;Sun, Jie;Zhu, Lin;Zhou, Jinzhu;Zhao, Yongxiang;Yu, Zhengyu;Sun, Bing;Guo, Rongli;He, Kongwang;Li, Bin;Li, Bin

作者机构：

关键词： PEDV; quantitative ELISA; antigen detection; intestinal and fecal samples; evaluation vaccine

期刊名称：FRONTIERS IN VETERINARY SCIENCE （ 影响因子：3.412； 五年影响因子：3.588 ）

ISSN：

年卷期： 2020 年 7 卷

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收录情况： SCI

摘要： Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in sucking piglets with a high mortality rate. Here, we developed a double antibody sandwich quantitative enzyme-linked immunosorbent assay (DAS-qELISA) for detection of PEDV using a specific monoclonal antibody against PEDV N protein and anti-PEDV rabbit serum. Using DAS-qELISA, the detection limit of recombinant PEDV N protein and virus titer were approximately 1 mu g/L and 10(2.0) TCID50/ml, respectively. A total of 90 intestinal and 237 fecal samples were then screened for the presence of PEDV using DAS-qELISA and reverse transcriptase PCR (RT-PCR). DAS-qELISA had a high specificity of 98.1% and sensitivity of 93.5%. The accuracy rate between DAS-qELISA and RT-PCR was 95.7%. More importantly, the viral antigen concentrations remained unchanged before and after one inactivated vaccine preparation by using the DAS-qELISA. These results suggest DAS-qELISA could be used for antigen detection of inactivated vaccine samples and clinical samples. It is a novel method for diagnosing diseases and evaluation of the PEDV vaccine.

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