Development and Validation of an Indirect and Blocking ELISA for the Serological Diagnosis of African Swine Fever
African swine fever (ASF) is an economically devastating viral disease of pigs caused by the ASF virus (ASFV). The rapid global spread of ASF has increased the demand for ASF diagnostics to be readily available and accessible. No commercial ASF enzyme-linked immunosorbent assay (ELISA) kits are manu...
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MDPI AG
2024-11-01
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| author | Chukwunonso Onyilagha Kaye Quizon Dmytro Zhmendak Ian El Kanoa Thang Truong Thanuja Ambagala Alfonso Clavijo Van Phan Le Shawn Babiuk Aruna Ambagala |
| author_facet | Chukwunonso Onyilagha Kaye Quizon Dmytro Zhmendak Ian El Kanoa Thang Truong Thanuja Ambagala Alfonso Clavijo Van Phan Le Shawn Babiuk Aruna Ambagala |
| author_sort | Chukwunonso Onyilagha |
| collection | DOAJ |
| description | African swine fever (ASF) is an economically devastating viral disease of pigs caused by the ASF virus (ASFV). The rapid global spread of ASF has increased the demand for ASF diagnostics to be readily available and accessible. No commercial ASF enzyme-linked immunosorbent assay (ELISA) kits are manufactured and licensed in North America. Here, we report the development of two serological diagnostic assays, a blocking ELISA (bELISA) based on ASFV glycoprotein p54 and an indirect ELISA (iELISA) based on ASFV glycoproteins p54 and p72. The assays showed high sensitivity and specificity and detected anti-ASFV antibodies in serum samples from experimentally infected animals as early as 8 days post-infection. The two assays were produced commercially (AsurDx<sup>™</sup> bELISA and iELISA) and subjected to extensive validation. Based on data from a set of characterized reference sera, the prototype commercial assays, while maintaining 100.00% specificity, showed 97.67% (AsurDx<sup>™</sup> bELISA) and 83.72% (AsurDx<sup>™</sup> iELISA) sensitivity. Both prototype assays detected anti-ASFV antibodies in serum samples collected from pigs experimentally infected with multiple ASFV strains and field samples collected from sick, recovering, and vaccinated animals. The two commercially available assays can be used in routine ASF diagnostics, serological surveys, and for evaluating serological responses to ASF vaccine candidates. |
| format | Article |
| id | doaj-art-0e3d67c7ed5a4ac3a2d06c2b3fa78b78 |
| institution | OA Journals |
| issn | 2076-0817 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj-art-0e3d67c7ed5a4ac3a2d06c2b3fa78b782025-08-20T02:04:55ZengMDPI AGPathogens2076-08172024-11-01131198110.3390/pathogens13110981Development and Validation of an Indirect and Blocking ELISA for the Serological Diagnosis of African Swine FeverChukwunonso Onyilagha0Kaye Quizon1Dmytro Zhmendak2Ian El Kanoa3Thang Truong4Thanuja Ambagala5Alfonso Clavijo6Van Phan Le7Shawn Babiuk8Aruna Ambagala9National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, CanadaNational Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB R3E 3M4, CanadaNational Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, CanadaNational Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, CanadaNational Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB R3E 3M4, CanadaNational Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, CanadaNational Bio and Agro-Defense Facility, Agricultural Research Service, United States Department of Agriculture, Manhattan, KS 66506, USADepartment of Microbiology and Infectious Disease, College of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi 100000, VietnamNational Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, CanadaNational Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, CanadaAfrican swine fever (ASF) is an economically devastating viral disease of pigs caused by the ASF virus (ASFV). The rapid global spread of ASF has increased the demand for ASF diagnostics to be readily available and accessible. No commercial ASF enzyme-linked immunosorbent assay (ELISA) kits are manufactured and licensed in North America. Here, we report the development of two serological diagnostic assays, a blocking ELISA (bELISA) based on ASFV glycoprotein p54 and an indirect ELISA (iELISA) based on ASFV glycoproteins p54 and p72. The assays showed high sensitivity and specificity and detected anti-ASFV antibodies in serum samples from experimentally infected animals as early as 8 days post-infection. The two assays were produced commercially (AsurDx<sup>™</sup> bELISA and iELISA) and subjected to extensive validation. Based on data from a set of characterized reference sera, the prototype commercial assays, while maintaining 100.00% specificity, showed 97.67% (AsurDx<sup>™</sup> bELISA) and 83.72% (AsurDx<sup>™</sup> iELISA) sensitivity. Both prototype assays detected anti-ASFV antibodies in serum samples collected from pigs experimentally infected with multiple ASFV strains and field samples collected from sick, recovering, and vaccinated animals. The two commercially available assays can be used in routine ASF diagnostics, serological surveys, and for evaluating serological responses to ASF vaccine candidates.https://www.mdpi.com/2076-0817/13/11/981African swine feverdiagnosticsindirect ELISAblocking ELISAserology |
| spellingShingle | Chukwunonso Onyilagha Kaye Quizon Dmytro Zhmendak Ian El Kanoa Thang Truong Thanuja Ambagala Alfonso Clavijo Van Phan Le Shawn Babiuk Aruna Ambagala Development and Validation of an Indirect and Blocking ELISA for the Serological Diagnosis of African Swine Fever Pathogens African swine fever diagnostics indirect ELISA blocking ELISA serology |
| title | Development and Validation of an Indirect and Blocking ELISA for the Serological Diagnosis of African Swine Fever |
| title_full | Development and Validation of an Indirect and Blocking ELISA for the Serological Diagnosis of African Swine Fever |
| title_fullStr | Development and Validation of an Indirect and Blocking ELISA for the Serological Diagnosis of African Swine Fever |
| title_full_unstemmed | Development and Validation of an Indirect and Blocking ELISA for the Serological Diagnosis of African Swine Fever |
| title_short | Development and Validation of an Indirect and Blocking ELISA for the Serological Diagnosis of African Swine Fever |
| title_sort | development and validation of an indirect and blocking elisa for the serological diagnosis of african swine fever |
| topic | African swine fever diagnostics indirect ELISA blocking ELISA serology |
| url | https://www.mdpi.com/2076-0817/13/11/981 |
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