Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virus
In recent years, African swine fever (ASF) has caused a devastating blow to the swine industry globally. Since no effective vaccine is available, strict biosafety measures and rapid diagnosis are the most effective strategies for ASF control. ASFV p30 is one of the most antigenic viral proteins that...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.988355/full |
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| author | Jingjing Ding Jifei Yang Daoyuan Jiang Yanyang Zhou Chenxi Li Chenxi Li Chenxi Li Yanhua Li Yanhua Li Yanhua Li |
| author_facet | Jingjing Ding Jifei Yang Daoyuan Jiang Yanyang Zhou Chenxi Li Chenxi Li Chenxi Li Yanhua Li Yanhua Li Yanhua Li |
| author_sort | Jingjing Ding |
| collection | DOAJ |
| description | In recent years, African swine fever (ASF) has caused a devastating blow to the swine industry globally. Since no effective vaccine is available, strict biosafety measures and rapid diagnosis are the most effective strategies for ASF control. ASFV p30 is one of the most antigenic viral proteins that have been widely used in the field for serological diagnosis of ASF infection. In this study, we developed a luciferase immunoprecipitation system (LIPS) assay for the detection of ASFV antibodies in pig serum using Gaussia luciferase (GLuc)-tagged p30 as a diagnostic antigen. The optimal GLuc-p30 input of 107 luminance units (LU) and optimal serum dilution factor of 1/100 were set to achieve the highest P/N ratio. Based on 87 ASFV-positive and negative pig sera, the cutoff value of the S/N ratio could be set between 2.298 and 30.59 to achieve 100% sensitivity and 100% specificity. Moreover, the diagnostic sensitivity of this LIPS is comparable to that of a commercial enzyme-linked immunosorbent assay (ELISA) and the specificity of LIPS is even superior to the tested ELISA. In conclusion, we have established a LIPS assay for ASFV antibody detection, which could be a potential method for ASFV diagnosis in laboratories and farms. |
| first_indexed | 2024-12-10T10:54:29Z |
| format | Article |
| id | doaj.art-282069d43dba4e8aa45b62bd0d72b482 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-12-10T10:54:29Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-282069d43dba4e8aa45b62bd0d72b4822022-12-22T01:51:54ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-09-011210.3389/fcimb.2022.988355988355Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virusJingjing Ding0Jifei Yang1Daoyuan Jiang2Yanyang Zhou3Chenxi Li4Chenxi Li5Chenxi Li6Yanhua Li7Yanhua Li8Yanhua Li9College of Veterinary Medicine, Yangzhou University, Yangzhou, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou, ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou, ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou, ChinaComparative Medicine Research Institute, Yangzhou University, Yangzhou, ChinaJiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou, ChinaComparative Medicine Research Institute, Yangzhou University, Yangzhou, ChinaJiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, ChinaIn recent years, African swine fever (ASF) has caused a devastating blow to the swine industry globally. Since no effective vaccine is available, strict biosafety measures and rapid diagnosis are the most effective strategies for ASF control. ASFV p30 is one of the most antigenic viral proteins that have been widely used in the field for serological diagnosis of ASF infection. In this study, we developed a luciferase immunoprecipitation system (LIPS) assay for the detection of ASFV antibodies in pig serum using Gaussia luciferase (GLuc)-tagged p30 as a diagnostic antigen. The optimal GLuc-p30 input of 107 luminance units (LU) and optimal serum dilution factor of 1/100 were set to achieve the highest P/N ratio. Based on 87 ASFV-positive and negative pig sera, the cutoff value of the S/N ratio could be set between 2.298 and 30.59 to achieve 100% sensitivity and 100% specificity. Moreover, the diagnostic sensitivity of this LIPS is comparable to that of a commercial enzyme-linked immunosorbent assay (ELISA) and the specificity of LIPS is even superior to the tested ELISA. In conclusion, we have established a LIPS assay for ASFV antibody detection, which could be a potential method for ASFV diagnosis in laboratories and farms.https://www.frontiersin.org/articles/10.3389/fcimb.2022.988355/fullAfrican swine fever virusluciferase immunoprecipitation systemgaussia luciferasep30p54Serological diagnosis |
| spellingShingle | Jingjing Ding Jifei Yang Daoyuan Jiang Yanyang Zhou Chenxi Li Chenxi Li Chenxi Li Yanhua Li Yanhua Li Yanhua Li Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virus Frontiers in Cellular and Infection Microbiology African swine fever virus luciferase immunoprecipitation system gaussia luciferase p30 p54 Serological diagnosis |
| title | Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virus |
| title_full | Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virus |
| title_fullStr | Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virus |
| title_full_unstemmed | Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virus |
| title_short | Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virus |
| title_sort | development of a highly sensitive gaussia luciferase immunoprecipitation assay for the detection of antibodies against african swine fever virus |
| topic | African swine fever virus luciferase immunoprecipitation system gaussia luciferase p30 p54 Serological diagnosis |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.988355/full |
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