Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen
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 PED...
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| Format: | Article |
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Frontiers Media S.A.
2020-10-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2020.540248/full |
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| author | Baochao Fan Baochao Fan Baochao Fan Baochao Fan Jie Sun Jie Sun Jie Sun Jie Sun Lin Zhu Lin Zhu Lin Zhu Lin Zhu Jinzhu Zhou Jinzhu Zhou Jinzhu Zhou Jinzhu Zhou Yongxiang Zhao Yongxiang Zhao Yongxiang Zhao Yongxiang Zhao Zhengyu Yu Zhengyu Yu Zhengyu Yu Zhengyu Yu Bing Sun Bing Sun Bing Sun Bing Sun Rongli Guo Rongli Guo Rongli Guo Rongli Guo Kongwang He Kongwang He Kongwang He Kongwang He Bin Li Bin Li Bin Li Bin Li Bin Li |
| author_facet | Baochao Fan Baochao Fan Baochao Fan Baochao Fan Jie Sun Jie Sun Jie Sun Jie Sun Lin Zhu Lin Zhu Lin Zhu Lin Zhu Jinzhu Zhou Jinzhu Zhou Jinzhu Zhou Jinzhu Zhou Yongxiang Zhao Yongxiang Zhao Yongxiang Zhao Yongxiang Zhao Zhengyu Yu Zhengyu Yu Zhengyu Yu Zhengyu Yu Bing Sun Bing Sun Bing Sun Bing Sun Rongli Guo Rongli Guo Rongli Guo Rongli Guo Kongwang He Kongwang He Kongwang He Kongwang He Bin Li Bin Li Bin Li Bin Li Bin Li |
| author_sort | Baochao Fan |
| collection | DOAJ |
| description | 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 μg/L and 102.0 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. |
| first_indexed | 2024-12-14T10:00:09Z |
| format | Article |
| id | doaj.art-280588ee53664c048bdaa16da02a5a46 |
| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-12-14T10:00:09Z |
| publishDate | 2020-10-01 |
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| series | Frontiers in Veterinary Science |
| spelling | doaj.art-280588ee53664c048bdaa16da02a5a462022-12-21T23:07:18ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692020-10-01710.3389/fvets.2020.540248540248Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus AntigenBaochao Fan0Baochao Fan1Baochao Fan2Baochao Fan3Jie Sun4Jie Sun5Jie Sun6Jie Sun7Lin Zhu8Lin Zhu9Lin Zhu10Lin Zhu11Jinzhu Zhou12Jinzhu Zhou13Jinzhu Zhou14Jinzhu Zhou15Yongxiang Zhao16Yongxiang Zhao17Yongxiang Zhao18Yongxiang Zhao19Zhengyu Yu20Zhengyu Yu21Zhengyu Yu22Zhengyu Yu23Bing Sun24Bing Sun25Bing Sun26Bing Sun27Rongli Guo28Rongli Guo29Rongli Guo30Rongli Guo31Kongwang He32Kongwang He33Kongwang He34Kongwang He35Bin Li36Bin Li37Bin Li38Bin Li39Bin Li40Key Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, ChinaJiangsu Co-infection Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, ChinaJiangsu Key Laboratory of Zoonoses, Yangzhou University, Yangzhou, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang, ChinaPorcine 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 μg/L and 102.0 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.https://www.frontiersin.org/articles/10.3389/fvets.2020.540248/fullPEDVquantitative ELISAantigen detectionintestinal and fecal samplesevaluation vaccine |
| spellingShingle | Baochao Fan Baochao Fan Baochao Fan Baochao Fan Jie Sun Jie Sun Jie Sun Jie Sun Lin Zhu Lin Zhu Lin Zhu Lin Zhu Jinzhu Zhou Jinzhu Zhou Jinzhu Zhou Jinzhu Zhou Yongxiang Zhao Yongxiang Zhao Yongxiang Zhao Yongxiang Zhao Zhengyu Yu Zhengyu Yu Zhengyu Yu Zhengyu Yu Bing Sun Bing Sun Bing Sun Bing Sun Rongli Guo Rongli Guo Rongli Guo Rongli Guo Kongwang He Kongwang He Kongwang He Kongwang He Bin Li Bin Li Bin Li Bin Li Bin Li Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen Frontiers in Veterinary Science PEDV quantitative ELISA antigen detection intestinal and fecal samples evaluation vaccine |
| title | Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen |
| title_full | Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen |
| title_fullStr | Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen |
| title_full_unstemmed | Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen |
| title_short | Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen |
| title_sort | development of a novel double antibody sandwich quantitative enzyme linked immunosorbent assay for detection of porcine epidemic diarrhea virus antigen |
| topic | PEDV quantitative ELISA antigen detection intestinal and fecal samples evaluation vaccine |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2020.540248/full |
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