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Development of a novel lateral flow assay for detection of African swine fever in blood

BACKGROUND: African swine fever (ASF) is a viral infectious disease of domestic and wild suids of all breeds and ages, causing a wide range of hemorrhagic syndromes and frequently characterized by high mortality. The disease is endemic in Sub-Saharan Africa and Sardinia. Since 2007, it has also been...

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Autores principales: Sastre, P., Gallardo, C., Monedero, A., Ruiz, T., Arias, M., Sanz, A., Rueda, P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5025629/
https://www.ncbi.nlm.nih.gov/pubmed/27633968
http://dx.doi.org/10.1186/s12917-016-0831-4
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author Sastre, P.
Gallardo, C.
Monedero, A.
Ruiz, T.
Arias, M.
Sanz, A.
Rueda, P.
author_facet Sastre, P.
Gallardo, C.
Monedero, A.
Ruiz, T.
Arias, M.
Sanz, A.
Rueda, P.
author_sort Sastre, P.
collection PubMed
description BACKGROUND: African swine fever (ASF) is a viral infectious disease of domestic and wild suids of all breeds and ages, causing a wide range of hemorrhagic syndromes and frequently characterized by high mortality. The disease is endemic in Sub-Saharan Africa and Sardinia. Since 2007, it has also been present in different countries of Eastern Europe, where control measures have not been effective so far. The continued spread poses a serious threat to the swine industry worldwide. In the absence of vaccine, early detection of infected animals is of paramount importance for control of the outbreak, to prevent the transmission of the virus to healthy animals and subsequent spreading of the disease. Current laboratory diagnosis is mainly based on virological methods (antigen and genome detection) and serodiagnosis. RESULTS: In the present work, a Lateral Flow Assay (LFA) for antigen detection has been developed and evaluated. The test is based on the use of a MAb against VP72 protein of ASFV, the major viral capsid protein and highly immunogenic. First experiments using VP72 viral and recombinant protein or inactivated culture virus showed promising results with a sensitivity similar to that of a commercially available Antigen-ELISA. Moreover, these strips were tested with blood from experimentally infected pigs and field animals and the results compared with those of PCR and Antigen-ELISA. For the experimentally infected samples, there was an excellent correlation between the LFA and the ELISA, while the PCR always showed to be more sensitive (38 % positive samples by PCR versus 27 % by LFA). The LFA was demonstrated to be positive for animals with circulating virus levels exceeding 10(4) HAU. With the field samples, once again, the PCR detected more positives than either the Antigen-ELISA or LFA, although here the number of positive samples scored by the LFA exceeded the values obtained with the Antigen-ELISA, showing 60 % positivity vs 48 % for the ELISA. For the two groups of sera, the specificity was close to 100 % indicating that hardly any false positive samples were found. CONCLUSIONS: The newly developed LFA allows rapid and reliable detection of ASFV, at field and laboratory level, providing a new useful tool for control programs and in situations where laboratory support and skilled personnel are limited.
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spelling pubmed-50256292016-09-20 Development of a novel lateral flow assay for detection of African swine fever in blood Sastre, P. Gallardo, C. Monedero, A. Ruiz, T. Arias, M. Sanz, A. Rueda, P. BMC Vet Res Methodology Article BACKGROUND: African swine fever (ASF) is a viral infectious disease of domestic and wild suids of all breeds and ages, causing a wide range of hemorrhagic syndromes and frequently characterized by high mortality. The disease is endemic in Sub-Saharan Africa and Sardinia. Since 2007, it has also been present in different countries of Eastern Europe, where control measures have not been effective so far. The continued spread poses a serious threat to the swine industry worldwide. In the absence of vaccine, early detection of infected animals is of paramount importance for control of the outbreak, to prevent the transmission of the virus to healthy animals and subsequent spreading of the disease. Current laboratory diagnosis is mainly based on virological methods (antigen and genome detection) and serodiagnosis. RESULTS: In the present work, a Lateral Flow Assay (LFA) for antigen detection has been developed and evaluated. The test is based on the use of a MAb against VP72 protein of ASFV, the major viral capsid protein and highly immunogenic. First experiments using VP72 viral and recombinant protein or inactivated culture virus showed promising results with a sensitivity similar to that of a commercially available Antigen-ELISA. Moreover, these strips were tested with blood from experimentally infected pigs and field animals and the results compared with those of PCR and Antigen-ELISA. For the experimentally infected samples, there was an excellent correlation between the LFA and the ELISA, while the PCR always showed to be more sensitive (38 % positive samples by PCR versus 27 % by LFA). The LFA was demonstrated to be positive for animals with circulating virus levels exceeding 10(4) HAU. With the field samples, once again, the PCR detected more positives than either the Antigen-ELISA or LFA, although here the number of positive samples scored by the LFA exceeded the values obtained with the Antigen-ELISA, showing 60 % positivity vs 48 % for the ELISA. For the two groups of sera, the specificity was close to 100 % indicating that hardly any false positive samples were found. CONCLUSIONS: The newly developed LFA allows rapid and reliable detection of ASFV, at field and laboratory level, providing a new useful tool for control programs and in situations where laboratory support and skilled personnel are limited. BioMed Central 2016-09-15 /pmc/articles/PMC5025629/ /pubmed/27633968 http://dx.doi.org/10.1186/s12917-016-0831-4 Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology Article
Sastre, P.
Gallardo, C.
Monedero, A.
Ruiz, T.
Arias, M.
Sanz, A.
Rueda, P.
Development of a novel lateral flow assay for detection of African swine fever in blood
title Development of a novel lateral flow assay for detection of African swine fever in blood
title_full Development of a novel lateral flow assay for detection of African swine fever in blood
title_fullStr Development of a novel lateral flow assay for detection of African swine fever in blood
title_full_unstemmed Development of a novel lateral flow assay for detection of African swine fever in blood
title_short Development of a novel lateral flow assay for detection of African swine fever in blood
title_sort development of a novel lateral flow assay for detection of african swine fever in blood
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5025629/
https://www.ncbi.nlm.nih.gov/pubmed/27633968
http://dx.doi.org/10.1186/s12917-016-0831-4
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