Cargando…
Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain
BACKGROUND: Porcine epidemic diarrhea (PED), a swine epidemic disease caused by porcine epidemic diarrhea virus (PEDV), is characterized by severe watery diarrhea, vomiting, dehydration and high mortality in piglets, and has caused serious economic losses to the global porcine industry. The level of...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9386190/ https://www.ncbi.nlm.nih.gov/pubmed/35982455 http://dx.doi.org/10.1186/s12917-022-03419-w |
_version_ | 1784769748276346880 |
---|---|
author | Wang, Kun Hu, Zhiqiang Fan, Mingyu Shao, Zhenwen Yu, Qiannan Li, Xiaowen |
author_facet | Wang, Kun Hu, Zhiqiang Fan, Mingyu Shao, Zhenwen Yu, Qiannan Li, Xiaowen |
author_sort | Wang, Kun |
collection | PubMed |
description | BACKGROUND: Porcine epidemic diarrhea (PED), a swine epidemic disease caused by porcine epidemic diarrhea virus (PEDV), is characterized by severe watery diarrhea, vomiting, dehydration and high mortality in piglets, and has caused serious economic losses to the global porcine industry. The level of PEDV IgA antibody is a key marker to assess the extent of passive immunity of the resistance against PEDV infection. However, current commercial structure proteins-based kits for detection of PEDV antibody are not affordable, and those kits require complicated antigen preparation procedures, which cannot meet the scope of economic benefits of many large-scale pig companies in China. Therefore, there is an urgent need to develop an accurate, simple, and economical method for IgA detection in clinical samples. In this study, an indirect ELISA (i-ELISA) method was developed based on a purified PEDV epidemic strain (NH-TA2020). RESULTS: The results show that optimal working dilution ratios of PEDV antigen and HRP anti-swine IgA are at 1: 1000 and 1:15000 respectively. The sensitivity of this method is high with the maximum dilution of samples up to 1:160, and coefficients of variation (CV) of both the intra assays and inter assays were no more than 15%. In addition, the relative sensitivities of the i-ELISA were above 90% compared with values from commercial kits in both serum and oral fluid samples. CONCLUSIONS: Our results suggested that the i-ELISA developed in this study was an accurate, simple, and economical method for PEDV-IgA detection in clinical samples. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-022-03419-w. |
format | Online Article Text |
id | pubmed-9386190 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-93861902022-08-18 Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain Wang, Kun Hu, Zhiqiang Fan, Mingyu Shao, Zhenwen Yu, Qiannan Li, Xiaowen BMC Vet Res Research BACKGROUND: Porcine epidemic diarrhea (PED), a swine epidemic disease caused by porcine epidemic diarrhea virus (PEDV), is characterized by severe watery diarrhea, vomiting, dehydration and high mortality in piglets, and has caused serious economic losses to the global porcine industry. The level of PEDV IgA antibody is a key marker to assess the extent of passive immunity of the resistance against PEDV infection. However, current commercial structure proteins-based kits for detection of PEDV antibody are not affordable, and those kits require complicated antigen preparation procedures, which cannot meet the scope of economic benefits of many large-scale pig companies in China. Therefore, there is an urgent need to develop an accurate, simple, and economical method for IgA detection in clinical samples. In this study, an indirect ELISA (i-ELISA) method was developed based on a purified PEDV epidemic strain (NH-TA2020). RESULTS: The results show that optimal working dilution ratios of PEDV antigen and HRP anti-swine IgA are at 1: 1000 and 1:15000 respectively. The sensitivity of this method is high with the maximum dilution of samples up to 1:160, and coefficients of variation (CV) of both the intra assays and inter assays were no more than 15%. In addition, the relative sensitivities of the i-ELISA were above 90% compared with values from commercial kits in both serum and oral fluid samples. CONCLUSIONS: Our results suggested that the i-ELISA developed in this study was an accurate, simple, and economical method for PEDV-IgA detection in clinical samples. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-022-03419-w. BioMed Central 2022-08-18 /pmc/articles/PMC9386190/ /pubmed/35982455 http://dx.doi.org/10.1186/s12917-022-03419-w Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Wang, Kun Hu, Zhiqiang Fan, Mingyu Shao, Zhenwen Yu, Qiannan Li, Xiaowen Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain |
title | Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain |
title_full | Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain |
title_fullStr | Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain |
title_full_unstemmed | Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain |
title_short | Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain |
title_sort | development of an indirect elisa to detect pedv specific iga antibody based on a pedv epidemic strain |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9386190/ https://www.ncbi.nlm.nih.gov/pubmed/35982455 http://dx.doi.org/10.1186/s12917-022-03419-w |
work_keys_str_mv | AT wangkun developmentofanindirectelisatodetectpedvspecificigaantibodybasedonapedvepidemicstrain AT huzhiqiang developmentofanindirectelisatodetectpedvspecificigaantibodybasedonapedvepidemicstrain AT fanmingyu developmentofanindirectelisatodetectpedvspecificigaantibodybasedonapedvepidemicstrain AT shaozhenwen developmentofanindirectelisatodetectpedvspecificigaantibodybasedonapedvepidemicstrain AT yuqiannan developmentofanindirectelisatodetectpedvspecificigaantibodybasedonapedvepidemicstrain AT lixiaowen developmentofanindirectelisatodetectpedvspecificigaantibodybasedonapedvepidemicstrain |