Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus
Epizootic haemorragic disease (EHD) is an important disease of white-tailed deer and can cause a bluetongue-like illness in cattle. A definitive diagnosis of EHD relies on molecular assays such as real-time RT-qPCR or conventional PCR. Reverse transcription loop-mediated isothermal amplification (RT...
| Autores principales: | , , , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
MDPI
2021
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/121008 |
| _version_ | 1855537706923196416 |
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| author | Rajko-Nenow, P. Howson, E.L.A. Clark, D. Hilton, N. Ambagala, A. Svitek, Nicholas Flannery, J. Batten, C. |
| author_browse | Ambagala, A. Batten, C. Clark, D. Flannery, J. Hilton, N. Howson, E.L.A. Rajko-Nenow, P. Svitek, Nicholas |
| author_facet | Rajko-Nenow, P. Howson, E.L.A. Clark, D. Hilton, N. Ambagala, A. Svitek, Nicholas Flannery, J. Batten, C. |
| author_sort | Rajko-Nenow, P. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Epizootic haemorragic disease (EHD) is an important disease of white-tailed deer and can cause a bluetongue-like illness in cattle. A definitive diagnosis of EHD relies on molecular assays such as real-time RT-qPCR or conventional PCR. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a cost-effective, specific, and sensitive technique that provides an alternative to RT-qPCR. We designed two sets of specific primers targeting segment-9 of the EHD virus genome to enable the detection of western and eastern topotypes, and evaluated their performance in singleplex and multiplex formats using cell culture isolates (n = 43), field specimens (n = 20), and a proficiency panel (n = 10). The limit of detection of the eastern and western RT-LAMP assays was estimated as ~24.36 CT and as ~29.37 CT in relation to real-time RT-qPCR, respectively, indicating a greater sensitivity of the western topotype singleplex RT-LAMP. The sensitivity of the western topotype RT-LAMP assay, relative to the RT-qPCR assay, was 72.2%, indicating that it could be theoretically used to detect viraemic cervines and bovines. For the first time, an RT-LAMP assay was developed for the rapid detection of the EHD virus that could be used as either a field test or high throughput screening tool in established laboratories to control the spread of EHD. |
| format | Journal Article |
| id | CGSpace121008 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | CGSpace1210082023-12-08T19:36:04Z Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus Rajko-Nenow, P. Howson, E.L.A. Clark, D. Hilton, N. Ambagala, A. Svitek, Nicholas Flannery, J. Batten, C. animal diseases diagnostic techniques Epizootic haemorragic disease (EHD) is an important disease of white-tailed deer and can cause a bluetongue-like illness in cattle. A definitive diagnosis of EHD relies on molecular assays such as real-time RT-qPCR or conventional PCR. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a cost-effective, specific, and sensitive technique that provides an alternative to RT-qPCR. We designed two sets of specific primers targeting segment-9 of the EHD virus genome to enable the detection of western and eastern topotypes, and evaluated their performance in singleplex and multiplex formats using cell culture isolates (n = 43), field specimens (n = 20), and a proficiency panel (n = 10). The limit of detection of the eastern and western RT-LAMP assays was estimated as ~24.36 CT and as ~29.37 CT in relation to real-time RT-qPCR, respectively, indicating a greater sensitivity of the western topotype singleplex RT-LAMP. The sensitivity of the western topotype RT-LAMP assay, relative to the RT-qPCR assay, was 72.2%, indicating that it could be theoretically used to detect viraemic cervines and bovines. For the first time, an RT-LAMP assay was developed for the rapid detection of the EHD virus that could be used as either a field test or high throughput screening tool in established laboratories to control the spread of EHD. 2021-11-15 2022-08-30T10:22:25Z 2022-08-30T10:22:25Z Journal Article https://hdl.handle.net/10568/121008 en Open Access MDPI Rajko-Nenow, P., Howson, E.L.A., Clark, D., Hilton, N., Ambagala, A., Svitek, N., Flannery, J. and Batten, C. 2021. Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus. Viruses 13(11): 2187. |
| spellingShingle | animal diseases diagnostic techniques Rajko-Nenow, P. Howson, E.L.A. Clark, D. Hilton, N. Ambagala, A. Svitek, Nicholas Flannery, J. Batten, C. Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus |
| title | Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus |
| title_full | Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus |
| title_fullStr | Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus |
| title_full_unstemmed | Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus |
| title_short | Development of a novel loop mediated isothermal amplification assay (LAMP) for the rapid detection of epizootic haemorrhagic disease virus |
| title_sort | development of a novel loop mediated isothermal amplification assay lamp for the rapid detection of epizootic haemorrhagic disease virus |
| topic | animal diseases diagnostic techniques |
| url | https://hdl.handle.net/10568/121008 |
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