Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA System
African swine fever (ASF) is a disease that affects both domestic and wild swine. It was recently reported in the Dominican Republic and Haiti (2021), representing a substantial risk to America. The goal of this study was to produce a truncated form of the ASF-p72 recombinant protein based on the AS...
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MDPI AG
2025-05-01
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| author | Julieta Sandra Cuevas-Romero Perla Lucero Zavala-Ocampo Sonia Pina-Pedrero Llilianne Ganges Adriana Muñoz-Aguilera José Bryan García-Cambrón Fernando Rodriguez Aruna Ambagala José Luis Cerriteño-Sánchez |
| author_facet | Julieta Sandra Cuevas-Romero Perla Lucero Zavala-Ocampo Sonia Pina-Pedrero Llilianne Ganges Adriana Muñoz-Aguilera José Bryan García-Cambrón Fernando Rodriguez Aruna Ambagala José Luis Cerriteño-Sánchez |
| author_sort | Julieta Sandra Cuevas-Romero |
| collection | DOAJ |
| description | African swine fever (ASF) is a disease that affects both domestic and wild swine. It was recently reported in the Dominican Republic and Haiti (2021), representing a substantial risk to America. The goal of this study was to produce a truncated form of the ASF-p72 recombinant protein based on the ASF strain genotype II (Georgia 2017) as well as to develop and validate a sensitive and specific ASF indirect-ELISA (iELISA) for early detection of ASF. The truncated ASF-p72 recombinant protein was successfully expressed in <i>E. coli</i> BL21/DE3 cells using the pET-SUMO plasmid. Bioinformatics analysis showed 100% homology among the new isolates of ASFV from genotype II. The ASF-p72-truncated protein was used to develop an iELISA, which had a high sensitivity (88%) and strong specificity (97%); the concordance index kappa was K = 0.872, indicating nearly perfect agreement compared to the WOAH confirmatory immunoperoxidase test. The validation results utilizing the reference sera panel from the OIE-ASF Reference Laboratory show the excellent detection capabilities of ASF antibodies up to a 1:1000 serum dilution. The inter-assay coefficient of variation (CV 10.4%) and intra-assay CV (2.8%) data show that the assay is precise and reproducible. This biotechnology advancement can be used to conduct future epidemiological research for ASF surveillance in ASF-free American countries. |
| format | Article |
| id | doaj-art-4de684d291ef4176b3c5fd97f0d7ca76 |
| institution | Kabale University |
| issn | 2076-0817 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
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| series | Pathogens |
| spelling | doaj-art-4de684d291ef4176b3c5fd97f0d7ca762025-08-20T03:27:40ZengMDPI AGPathogens2076-08172025-05-0114654210.3390/pathogens14060542Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA SystemJulieta Sandra Cuevas-Romero0Perla Lucero Zavala-Ocampo1Sonia Pina-Pedrero2Llilianne Ganges3Adriana Muñoz-Aguilera4José Bryan García-Cambrón5Fernando Rodriguez6Aruna Ambagala7José Luis Cerriteño-Sánchez8Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Cuajimalpa de Morelos, Mexico City 05110, MexicoPosgrado en Ciencias de la Producción y de la Salud Animal, Facultad de Medicina Veterinaria, Universidad Nacional Autónoma de México. Av. Universidad 3004, Copilco Universidad, Mexico City 04510, MexicoIRTA, Animal Health, Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), 08193 Bellaterra, SpainIRTA, Animal Health, Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), 08193 Bellaterra, SpainIRTA, Animal Health, Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), 08193 Bellaterra, SpainPosgrado en Biología Experimental, Universidad Autónoma Metropolitana Iztapalapa, Mexico City 09340, MexicoIRTA, Animal Health, Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), 08193 Bellaterra, SpainNational Centre for Foreign Animal Disease, Winnipeg, MB R3E 3M4, CanadaCentro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Cuajimalpa de Morelos, Mexico City 05110, MexicoAfrican swine fever (ASF) is a disease that affects both domestic and wild swine. It was recently reported in the Dominican Republic and Haiti (2021), representing a substantial risk to America. The goal of this study was to produce a truncated form of the ASF-p72 recombinant protein based on the ASF strain genotype II (Georgia 2017) as well as to develop and validate a sensitive and specific ASF indirect-ELISA (iELISA) for early detection of ASF. The truncated ASF-p72 recombinant protein was successfully expressed in <i>E. coli</i> BL21/DE3 cells using the pET-SUMO plasmid. Bioinformatics analysis showed 100% homology among the new isolates of ASFV from genotype II. The ASF-p72-truncated protein was used to develop an iELISA, which had a high sensitivity (88%) and strong specificity (97%); the concordance index kappa was K = 0.872, indicating nearly perfect agreement compared to the WOAH confirmatory immunoperoxidase test. The validation results utilizing the reference sera panel from the OIE-ASF Reference Laboratory show the excellent detection capabilities of ASF antibodies up to a 1:1000 serum dilution. The inter-assay coefficient of variation (CV 10.4%) and intra-assay CV (2.8%) data show that the assay is precise and reproducible. This biotechnology advancement can be used to conduct future epidemiological research for ASF surveillance in ASF-free American countries.https://www.mdpi.com/2076-0817/14/6/542p72African swine feverrecombinant protein |
| spellingShingle | Julieta Sandra Cuevas-Romero Perla Lucero Zavala-Ocampo Sonia Pina-Pedrero Llilianne Ganges Adriana Muñoz-Aguilera José Bryan García-Cambrón Fernando Rodriguez Aruna Ambagala José Luis Cerriteño-Sánchez Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA System Pathogens p72 African swine fever recombinant protein |
| title | Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA System |
| title_full | Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA System |
| title_fullStr | Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA System |
| title_full_unstemmed | Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA System |
| title_short | Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA System |
| title_sort | cloning and expression of a truncated form of the p72 protein of the african swine fever virus asfv for application in an efficient indirect elisa system |
| topic | p72 African swine fever recombinant protein |
| url | https://www.mdpi.com/2076-0817/14/6/542 |
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