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Development Real-Time PCR Assays to Genetically Differentiate Vaccinated Pigs From Infected Pigs With the Eurasian Strain of African Swine Fever Virus

Currently, African swine fever virus (ASFV) represents one of the most important economic threats for the global pork industry. Recently, significant advances have been made in the development of potential vaccine candidates to protect pigs against this virus. We have previously developed attenuated...

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Autores principales: Velazquez-Salinas, Lauro, Ramirez-Medina, Elizabeth, Rai, Ayushi, Pruitt, Sarah, Vuono, Elizabeth A., Espinoza, Nallely, Gladue, Douglas P., Borca, Manuel V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8579032/
https://www.ncbi.nlm.nih.gov/pubmed/34778441
http://dx.doi.org/10.3389/fvets.2021.768869
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author Velazquez-Salinas, Lauro
Ramirez-Medina, Elizabeth
Rai, Ayushi
Pruitt, Sarah
Vuono, Elizabeth A.
Espinoza, Nallely
Gladue, Douglas P.
Borca, Manuel V.
author_facet Velazquez-Salinas, Lauro
Ramirez-Medina, Elizabeth
Rai, Ayushi
Pruitt, Sarah
Vuono, Elizabeth A.
Espinoza, Nallely
Gladue, Douglas P.
Borca, Manuel V.
author_sort Velazquez-Salinas, Lauro
collection PubMed
description Currently, African swine fever virus (ASFV) represents one of the most important economic threats for the global pork industry. Recently, significant advances have been made in the development of potential vaccine candidates to protect pigs against this virus. We have previously developed attenuated vaccine candidates by deleting critical viral genes associated with virulence. Here, we present the development of the accompanying genetic tests to discriminate between infected and vaccinated animals (DIVA), a necessity during an ASFV vaccination campaign. We describe here the development of three independent real-time polymerase chain reaction (qPCR) assays that detect the presence of MGF-360-12L, UK, and I177L genes, which were previously deleted from the highly virulent Georgia strain of ASFV to produce the three recombinant live attenuated vaccine candidates. When compared with the diagnostic reference qPCR that detects the p72 gene, all assays demonstrated comparable levels of sensitivity, specificity, and efficiency of amplification to detect presence/absence of the ASFV Georgia 2007/1 strain (prototype virus of the Eurasian lineage) from a panel of blood samples from naïve, vaccinated, and infected pigs. Collectively, the results of this study demonstrate the potential of these real-time PCR assays to be used as genetic DIVA tests, supporting vaccination campaigns associated with the use of ASFV-ΔMGF, ASFV-G-Δ9GL/ΔUK, and ASFV-ΔI177L or cell culture adapted ASFV-ΔI177LΔLVR live attenuated vaccines in the field.
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spelling pubmed-85790322021-11-11 Development Real-Time PCR Assays to Genetically Differentiate Vaccinated Pigs From Infected Pigs With the Eurasian Strain of African Swine Fever Virus Velazquez-Salinas, Lauro Ramirez-Medina, Elizabeth Rai, Ayushi Pruitt, Sarah Vuono, Elizabeth A. Espinoza, Nallely Gladue, Douglas P. Borca, Manuel V. Front Vet Sci Veterinary Science Currently, African swine fever virus (ASFV) represents one of the most important economic threats for the global pork industry. Recently, significant advances have been made in the development of potential vaccine candidates to protect pigs against this virus. We have previously developed attenuated vaccine candidates by deleting critical viral genes associated with virulence. Here, we present the development of the accompanying genetic tests to discriminate between infected and vaccinated animals (DIVA), a necessity during an ASFV vaccination campaign. We describe here the development of three independent real-time polymerase chain reaction (qPCR) assays that detect the presence of MGF-360-12L, UK, and I177L genes, which were previously deleted from the highly virulent Georgia strain of ASFV to produce the three recombinant live attenuated vaccine candidates. When compared with the diagnostic reference qPCR that detects the p72 gene, all assays demonstrated comparable levels of sensitivity, specificity, and efficiency of amplification to detect presence/absence of the ASFV Georgia 2007/1 strain (prototype virus of the Eurasian lineage) from a panel of blood samples from naïve, vaccinated, and infected pigs. Collectively, the results of this study demonstrate the potential of these real-time PCR assays to be used as genetic DIVA tests, supporting vaccination campaigns associated with the use of ASFV-ΔMGF, ASFV-G-Δ9GL/ΔUK, and ASFV-ΔI177L or cell culture adapted ASFV-ΔI177LΔLVR live attenuated vaccines in the field. Frontiers Media S.A. 2021-10-27 /pmc/articles/PMC8579032/ /pubmed/34778441 http://dx.doi.org/10.3389/fvets.2021.768869 Text en Copyright © 2021 Velazquez-Salinas, Ramirez-Medina, Rai, Pruitt, Vuono, Espinoza, Gladue and Borca. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Velazquez-Salinas, Lauro
Ramirez-Medina, Elizabeth
Rai, Ayushi
Pruitt, Sarah
Vuono, Elizabeth A.
Espinoza, Nallely
Gladue, Douglas P.
Borca, Manuel V.
Development Real-Time PCR Assays to Genetically Differentiate Vaccinated Pigs From Infected Pigs With the Eurasian Strain of African Swine Fever Virus
title Development Real-Time PCR Assays to Genetically Differentiate Vaccinated Pigs From Infected Pigs With the Eurasian Strain of African Swine Fever Virus
title_full Development Real-Time PCR Assays to Genetically Differentiate Vaccinated Pigs From Infected Pigs With the Eurasian Strain of African Swine Fever Virus
title_fullStr Development Real-Time PCR Assays to Genetically Differentiate Vaccinated Pigs From Infected Pigs With the Eurasian Strain of African Swine Fever Virus
title_full_unstemmed Development Real-Time PCR Assays to Genetically Differentiate Vaccinated Pigs From Infected Pigs With the Eurasian Strain of African Swine Fever Virus
title_short Development Real-Time PCR Assays to Genetically Differentiate Vaccinated Pigs From Infected Pigs With the Eurasian Strain of African Swine Fever Virus
title_sort development real-time pcr assays to genetically differentiate vaccinated pigs from infected pigs with the eurasian strain of african swine fever virus
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8579032/
https://www.ncbi.nlm.nih.gov/pubmed/34778441
http://dx.doi.org/10.3389/fvets.2021.768869
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