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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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Autores principales: Fan, Baochao, Sun, Jie, Zhu, Lin, Zhou, Jinzhu, Zhao, Yongxiang, Yu, Zhengyu, Sun, Bing, Guo, Rongli, He, Kongwang, Li, Bin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7649156/
https://www.ncbi.nlm.nih.gov/pubmed/33195513
http://dx.doi.org/10.3389/fvets.2020.540248
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author Fan, Baochao
Sun, Jie
Zhu, Lin
Zhou, Jinzhu
Zhao, Yongxiang
Yu, Zhengyu
Sun, Bing
Guo, Rongli
He, Kongwang
Li, Bin
author_facet Fan, Baochao
Sun, Jie
Zhu, Lin
Zhou, Jinzhu
Zhao, Yongxiang
Yu, Zhengyu
Sun, Bing
Guo, Rongli
He, Kongwang
Li, Bin
author_sort Fan, Baochao
collection PubMed
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 10(2.0) TCID(50)/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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spelling pubmed-76491562020-11-13 Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen Fan, Baochao Sun, Jie Zhu, Lin Zhou, Jinzhu Zhao, Yongxiang Yu, Zhengyu Sun, Bing Guo, Rongli He, Kongwang Li, Bin Front Vet Sci Veterinary Science 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 10(2.0) TCID(50)/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. Frontiers Media S.A. 2020-10-26 /pmc/articles/PMC7649156/ /pubmed/33195513 http://dx.doi.org/10.3389/fvets.2020.540248 Text en Copyright © 2020 Fan, Sun, Zhu, Zhou, Zhao, Yu, Sun, Guo, He and Li. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Fan, Baochao
Sun, Jie
Zhu, Lin
Zhou, Jinzhu
Zhao, Yongxiang
Yu, Zhengyu
Sun, Bing
Guo, Rongli
He, Kongwang
Li, Bin
Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen
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 Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7649156/
https://www.ncbi.nlm.nih.gov/pubmed/33195513
http://dx.doi.org/10.3389/fvets.2020.540248
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