Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against African Swine Fever Virus
The incursion of African swine fever virus (ASFV) into Eurasia presents a threat to the world’s swine industry. Highly sensitive and specific diagnostic assays are urgently needed for rapid detection during an outbreak, post-outbreak investigation, and disease surveillance. In this study, a highly s...
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MDPI AG
2021-06-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/6/760 |
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| author | Fangfeng Yuan Vlad Petrovan Luis Gabriel Gimenez-Lirola Jeffrey J. Zimmerman Raymond R. R. Rowland Ying Fang |
| author_facet | Fangfeng Yuan Vlad Petrovan Luis Gabriel Gimenez-Lirola Jeffrey J. Zimmerman Raymond R. R. Rowland Ying Fang |
| author_sort | Fangfeng Yuan |
| collection | DOAJ |
| description | The incursion of African swine fever virus (ASFV) into Eurasia presents a threat to the world’s swine industry. Highly sensitive and specific diagnostic assays are urgently needed for rapid detection during an outbreak, post-outbreak investigation, and disease surveillance. In this study, a highly specific and repeatable blocking ELISA (bELISA) was developed using a recombinant p30 protein as the antigen combined with biotinylated mAb against p30 as the detection antibody. Initial test validation included sera from 810 uninfected animals and 106 animals experimentally inoculated with ASFV or recombinant alphavirus/adenovirus expressing p30. Receiver operating characteristic (ROC) analysis of the data calculated an optimal percentage of inhibition (PI) cutoff value of 45.92%, giving a diagnostic sensitivity of 98.11% and diagnostic specificity of 99.42%. The coefficient of variation of an internal quality control serum was 6.81% for between runs, 6.71% for within run, and 6.14% for within plate. A time course study of infected pigs showed that bELISA was able to detect seroconversion as early as 7 days post-inoculation. Taken together, these results demonstrate that bELISA can be used as an alternative serological test for detecting ASFV infection. |
| first_indexed | 2024-03-10T10:20:19Z |
| format | Article |
| id | doaj.art-4522672d22ed41aa88203543c65ff355 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T10:20:19Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-4522672d22ed41aa88203543c65ff3552023-11-22T00:26:59ZengMDPI AGPathogens2076-08172021-06-0110676010.3390/pathogens10060760Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against African Swine Fever VirusFangfeng Yuan0Vlad Petrovan1Luis Gabriel Gimenez-Lirola2Jeffrey J. Zimmerman3Raymond R. R. Rowland4Ying Fang5Department of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana Champaign, Urbana, IL 61802, USADepartment of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, USADepartment of Veterinary Diagnostic & Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USADepartment of Veterinary Diagnostic & Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USADepartment of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana Champaign, Urbana, IL 61802, USADepartment of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana Champaign, Urbana, IL 61802, USAThe incursion of African swine fever virus (ASFV) into Eurasia presents a threat to the world’s swine industry. Highly sensitive and specific diagnostic assays are urgently needed for rapid detection during an outbreak, post-outbreak investigation, and disease surveillance. In this study, a highly specific and repeatable blocking ELISA (bELISA) was developed using a recombinant p30 protein as the antigen combined with biotinylated mAb against p30 as the detection antibody. Initial test validation included sera from 810 uninfected animals and 106 animals experimentally inoculated with ASFV or recombinant alphavirus/adenovirus expressing p30. Receiver operating characteristic (ROC) analysis of the data calculated an optimal percentage of inhibition (PI) cutoff value of 45.92%, giving a diagnostic sensitivity of 98.11% and diagnostic specificity of 99.42%. The coefficient of variation of an internal quality control serum was 6.81% for between runs, 6.71% for within run, and 6.14% for within plate. A time course study of infected pigs showed that bELISA was able to detect seroconversion as early as 7 days post-inoculation. Taken together, these results demonstrate that bELISA can be used as an alternative serological test for detecting ASFV infection.https://www.mdpi.com/2076-0817/10/6/760African swine fever virusASFV monoclonal antibodyp30blocking ELISA |
| spellingShingle | Fangfeng Yuan Vlad Petrovan Luis Gabriel Gimenez-Lirola Jeffrey J. Zimmerman Raymond R. R. Rowland Ying Fang Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against African Swine Fever Virus Pathogens African swine fever virus ASFV monoclonal antibody p30 blocking ELISA |
| title | Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against African Swine Fever Virus |
| title_full | Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against African Swine Fever Virus |
| title_fullStr | Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against African Swine Fever Virus |
| title_full_unstemmed | Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against African Swine Fever Virus |
| title_short | Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against African Swine Fever Virus |
| title_sort | development of a blocking enzyme linked immunosorbent assay for detection of antibodies against african swine fever virus |
| topic | African swine fever virus ASFV monoclonal antibody p30 blocking ELISA |
| url | https://www.mdpi.com/2076-0817/10/6/760 |
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