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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

作者： Fan, Baochao 1 ; Sun, Jie 1 ; Zhu, Lin 1 ; Zhou, Jinzhu 1 ; Zhao, Yongxiang 1 ; Yu, Zhengyu 1 ; Sun, Bing 1 ; Guo, Rongli 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Med, Minist Agr, Key Lab Vet Biol Engn & Technol, Nanjing, Peoples R China

2.Minist Sci & Technol, State Key Lab Cultivat Base, Jiangsu Key Lab Food Qual & Safety, Nanjing, Peoples R China

3.Jiangsu Coinfect Ctr Prevent & Control Important, Yangzhou, Jiangsu, Peoples R China

4.Yangzhou Univ, Jiangsu Key Lab Zoonoses, Yangzhou, Jiangsu, Peoples R China

5.Jiangsu Univ, Sch Food & Biol Engn, Zhenjiang, Jiangsu, Peoples R China

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

页码：

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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