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Open AccessArticle

Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste

1
Agriculture Victoria Research, AgriBio Centre for AgriBioscience, Bundoora, VIC 3083, Australia
2
Ministry of Agriculture and Fisheries, Government of Timor-Leste, Av. Nicolao Lobato, Comoro, Dili 0332, Timor-Leste
3
Agriculture Victoria, Biosecurity and Agriculture Services, Bairnsdale, VIC 3857, Australia
*
Author to whom correspondence should be addressed.
Academic Editors: Douglas Gladue, Fernando Rodriguez and Manuel Borca
Viruses 2020, 12(12), 1444; https://doi.org/10.3390/v12121444
Received: 7 December 2020 / Accepted: 11 December 2020 / Published: 15 December 2020
(This article belongs to the Special Issue Endemic and Emerging Swine Viruses)
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. View Full-Text
Keywords: ASFV; LAMP; qPCR; colourmetric LAMP; Timor-Leste ASFV; LAMP; qPCR; colourmetric LAMP; Timor-Leste
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MDPI and ACS Style

Mee, P.T.; Wong, S.; O’Riley, K.J.; da Conceição, F.; Bendita da Costa Jong, J.; Phillips, D.E.; Rodoni, B.C.; Rawlin, G.T.; Lynch, S.E. Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste. Viruses 2020, 12, 1444. https://doi.org/10.3390/v12121444

AMA Style

Mee PT, Wong S, O’Riley KJ, da Conceição F, Bendita da Costa Jong J, Phillips DE, Rodoni BC, Rawlin GT, Lynch SE. Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste. Viruses. 2020; 12(12):1444. https://doi.org/10.3390/v12121444

Chicago/Turabian Style

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. 2020. "Field Verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) Assay during an Outbreak in Timor-Leste" Viruses 12, no. 12: 1444. https://doi.org/10.3390/v12121444

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