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Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus
BACKGROUND: Lumpy skin disease virus (LSDV) is a Capripoxvirus infecting cattle and Buffalos. Lumpy skin disease (LSD) leads to significant economic losses due to hide damage, reduction of milk production, mastitis, infertility and mortalities (10 %). Early detection of the virus is crucial to start...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5094145/ https://www.ncbi.nlm.nih.gov/pubmed/27806722 http://dx.doi.org/10.1186/s12917-016-0875-5 |
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author | Shalaby, Mohamed A. El-Deeb, Ayman El-Tholoth, Mohamed Hoffmann, Donata Czerny, Claus-Peter Hufert, Frank T. Weidmann, Manfred Abd El Wahed, Ahmed |
author_facet | Shalaby, Mohamed A. El-Deeb, Ayman El-Tholoth, Mohamed Hoffmann, Donata Czerny, Claus-Peter Hufert, Frank T. Weidmann, Manfred Abd El Wahed, Ahmed |
author_sort | Shalaby, Mohamed A. |
collection | PubMed |
description | BACKGROUND: Lumpy skin disease virus (LSDV) is a Capripoxvirus infecting cattle and Buffalos. Lumpy skin disease (LSD) leads to significant economic losses due to hide damage, reduction of milk production, mastitis, infertility and mortalities (10 %). Early detection of the virus is crucial to start appropriate outbreak control measures. Veterinarians rely on the presence of the characteristic clinical signs of LSD. Laboratory diagnostics including virus isolation, sequencing and real-time polymerase chain reaction (PCR) are performed at well-equipped laboratories. In this study, a portable, simple, and rapid recombinase polymerase amplification (RPA) assay for the detection of LSDV-genome for the use on farms was developed. RESULTS: The LSDV RPA assay was performed at 42 °C and detected down to 179 DNA copies/reaction in a maximum of 15 min. Unspecific amplification was observed with neither LSDV-negative samples (n = 12) nor nucleic acid preparations from orf virus, bovine papular stomatitis virus, cowpoxvirus, Peste des petits ruminants and Blue tongue virus (serotypes 1, 6 and 8). The clinical sensitivity of the LSDV RPA assay matched 100 % (n = 22) to real-time PCR results. In addition, the LSDV RPA assay detected sheep and goat poxviruses. CONCLUSION: The LSDV RPA assay is a rapid and sensitive test that could be implemented in field or at quarantine stations for the identification of LSDV infected case. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12917-016-0875-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5094145 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-50941452016-11-07 Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus Shalaby, Mohamed A. El-Deeb, Ayman El-Tholoth, Mohamed Hoffmann, Donata Czerny, Claus-Peter Hufert, Frank T. Weidmann, Manfred Abd El Wahed, Ahmed BMC Vet Res Methodology Article BACKGROUND: Lumpy skin disease virus (LSDV) is a Capripoxvirus infecting cattle and Buffalos. Lumpy skin disease (LSD) leads to significant economic losses due to hide damage, reduction of milk production, mastitis, infertility and mortalities (10 %). Early detection of the virus is crucial to start appropriate outbreak control measures. Veterinarians rely on the presence of the characteristic clinical signs of LSD. Laboratory diagnostics including virus isolation, sequencing and real-time polymerase chain reaction (PCR) are performed at well-equipped laboratories. In this study, a portable, simple, and rapid recombinase polymerase amplification (RPA) assay for the detection of LSDV-genome for the use on farms was developed. RESULTS: The LSDV RPA assay was performed at 42 °C and detected down to 179 DNA copies/reaction in a maximum of 15 min. Unspecific amplification was observed with neither LSDV-negative samples (n = 12) nor nucleic acid preparations from orf virus, bovine papular stomatitis virus, cowpoxvirus, Peste des petits ruminants and Blue tongue virus (serotypes 1, 6 and 8). The clinical sensitivity of the LSDV RPA assay matched 100 % (n = 22) to real-time PCR results. In addition, the LSDV RPA assay detected sheep and goat poxviruses. CONCLUSION: The LSDV RPA assay is a rapid and sensitive test that could be implemented in field or at quarantine stations for the identification of LSDV infected case. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12917-016-0875-5) contains supplementary material, which is available to authorized users. BioMed Central 2016-11-02 /pmc/articles/PMC5094145/ /pubmed/27806722 http://dx.doi.org/10.1186/s12917-016-0875-5 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 Shalaby, Mohamed A. El-Deeb, Ayman El-Tholoth, Mohamed Hoffmann, Donata Czerny, Claus-Peter Hufert, Frank T. Weidmann, Manfred Abd El Wahed, Ahmed Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus |
title | Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus |
title_full | Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus |
title_fullStr | Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus |
title_full_unstemmed | Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus |
title_short | Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus |
title_sort | recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5094145/ https://www.ncbi.nlm.nih.gov/pubmed/27806722 http://dx.doi.org/10.1186/s12917-016-0875-5 |
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