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Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste
Recent outbreaks of African swine fever virus (ASFV) have seen the movement of this virus into multiple new regions with devastating impact. Many of these outbreaks are occurring in remote, or resource-limited areas, that do not have access to molecular laboratories. Loop-mediated isothermal amplifi...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7765541/ https://www.ncbi.nlm.nih.gov/pubmed/33334037 http://dx.doi.org/10.3390/v12121444 |
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author | Mee, Peter T. Wong, Shani O’Riley, Kim J. da Conceição, Felisiano Bendita da Costa Jong, Joanita Phillips, Dianne E. Rodoni, Brendan C. Rawlin, Grant T. Lynch, Stacey E. |
author_facet | Mee, Peter T. Wong, Shani O’Riley, Kim J. da Conceição, Felisiano Bendita da Costa Jong, Joanita Phillips, Dianne E. Rodoni, Brendan C. Rawlin, Grant T. Lynch, Stacey E. |
author_sort | Mee, Peter T. |
collection | PubMed |
description | Recent outbreaks of African swine fever virus (ASFV) have seen the movement of this virus into multiple new regions with devastating impact. Many of these outbreaks are occurring in remote, or resource-limited areas, that do not have access to molecular laboratories. Loop-mediated isothermal amplification (LAMP) is a rapid point of care test that can overcome a range of inhibitors. We outline further development of a real-time ASFV LAMP, including field verification during an outbreak in Timor-Leste. To increase field applicability, the extraction step was removed and an internal amplification control (IAC) was implemented. Assay performance was assessed in six different sample matrices and verified for a range of clinical samples. A LAMP detection limit of 400 copies/rxn was determined based on synthetic positive control spikes. A colourmetric LAMP assay was also assessed on serum samples. Comparison of the LAMP assay to a quantitative polymerase chain reaction (qPCR) was performed on clinical ASFV samples, using both serum and oral/rectal swabs, with a substantial level of agreement observed. The further verification of the ASFV LAMP assay, removal of extraction step, implementation of an IAC and the assessment of a range of sample matrix, further support the use of this assay for rapid in-field detection of ASFV. |
format | Online Article Text |
id | pubmed-7765541 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-77655412020-12-27 Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste Mee, Peter T. Wong, Shani O’Riley, Kim J. da Conceição, Felisiano Bendita da Costa Jong, Joanita Phillips, Dianne E. Rodoni, Brendan C. Rawlin, Grant T. Lynch, Stacey E. Viruses Article Recent outbreaks of African swine fever virus (ASFV) have seen the movement of this virus into multiple new regions with devastating impact. Many of these outbreaks are occurring in remote, or resource-limited areas, that do not have access to molecular laboratories. Loop-mediated isothermal amplification (LAMP) is a rapid point of care test that can overcome a range of inhibitors. We outline further development of a real-time ASFV LAMP, including field verification during an outbreak in Timor-Leste. To increase field applicability, the extraction step was removed and an internal amplification control (IAC) was implemented. Assay performance was assessed in six different sample matrices and verified for a range of clinical samples. A LAMP detection limit of 400 copies/rxn was determined based on synthetic positive control spikes. A colourmetric LAMP assay was also assessed on serum samples. Comparison of the LAMP assay to a quantitative polymerase chain reaction (qPCR) was performed on clinical ASFV samples, using both serum and oral/rectal swabs, with a substantial level of agreement observed. The further verification of the ASFV LAMP assay, removal of extraction step, implementation of an IAC and the assessment of a range of sample matrix, further support the use of this assay for rapid in-field detection of ASFV. MDPI 2020-12-15 /pmc/articles/PMC7765541/ /pubmed/33334037 http://dx.doi.org/10.3390/v12121444 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Mee, Peter T. Wong, Shani O’Riley, Kim J. da Conceição, Felisiano Bendita da Costa Jong, Joanita Phillips, Dianne E. Rodoni, Brendan C. Rawlin, Grant T. Lynch, Stacey E. Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste |
title | Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste |
title_full | Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste |
title_fullStr | Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste |
title_full_unstemmed | Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste |
title_short | Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste |
title_sort | field verification of an african swine fever virus loop-mediated isothermal amplification (lamp) assay during an outbreak in timor-leste |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7765541/ https://www.ncbi.nlm.nih.gov/pubmed/33334037 http://dx.doi.org/10.3390/v12121444 |
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