Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain
Abstract 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...
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| Format: | Article |
| Language: | English |
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BMC
2022-08-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-022-03419-w |
| _version_ | 1798038900088766464 |
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| author | Kun Wang Zhiqiang Hu Mingyu Fan Zhenwen Shao Qiannan Yu Xiaowen Li |
| author_facet | Kun Wang Zhiqiang Hu Mingyu Fan Zhenwen Shao Qiannan Yu Xiaowen Li |
| author_sort | Kun Wang |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-04-11T21:46:40Z |
| format | Article |
| id | doaj.art-5cf05b326b0c427c81759f74ea7a5f00 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-04-11T21:46:40Z |
| publishDate | 2022-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-5cf05b326b0c427c81759f74ea7a5f002022-12-22T04:01:23ZengBMCBMC Veterinary Research1746-61482022-08-011811910.1186/s12917-022-03419-wDevelopment of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strainKun Wang0Zhiqiang Hu1Mingyu Fan2Zhenwen Shao3Qiannan Yu4Xiaowen Li5Shandong New Hope Liuhe Agriculture and Animal Husbandry Technology Co., Ltd (NHLH Academy of Swine Research)Shandong New Hope Liuhe Agriculture and Animal Husbandry Technology Co., Ltd (NHLH Academy of Swine Research)Shandong New Hope Liuhe Agriculture and Animal Husbandry Technology Co., Ltd (NHLH Academy of Swine Research)Shandong New Hope Liuhe Agriculture and Animal Husbandry Technology Co., Ltd (NHLH Academy of Swine Research)Shandong New Hope Liuhe Agriculture and Animal Husbandry Technology Co., Ltd (NHLH Academy of Swine Research)Shandong New Hope Liuhe Agriculture and Animal Husbandry Technology Co., Ltd (NHLH Academy of Swine Research)Abstract 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.https://doi.org/10.1186/s12917-022-03419-wPEDVNH-TA2020Indirect ELISAIgAIDEXX |
| spellingShingle | Kun Wang Zhiqiang Hu Mingyu Fan Zhenwen Shao Qiannan Yu Xiaowen Li Development of an indirect ELISA to detect PEDV specific IgA antibody based on a PEDV epidemic strain BMC Veterinary Research PEDV NH-TA2020 Indirect ELISA IgA IDEXX |
| 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 | PEDV NH-TA2020 Indirect ELISA IgA IDEXX |
| url | https://doi.org/10.1186/s12917-022-03419-w |
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