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Field verification of an African Swine Fever Virus Loop-Mediated Isothermal Amplification (LAMP) assay during an outbreak in Timor-Leste

journal contribution
posted on 04.01.2021, 05:05 by Peter T Mee, Shani Wong, Kim J O'Riley, Felisiano da Conceição, Joanita Bendita da Costa Jong, Dianne E Phillips, Brendan Rodoni, Grant Rawlin, Stacey E Lynch
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.

Funding

This research was funded by the Department of Agriculture grant number B8FV69A, the Department of Foreign Affairs and Trade grant number 106459 and Agriculture Victoria Research.

History

Publication Date

15/12/2020

Journal

Viruses

Volume

12

Issue

12

Article Number

1444

Pagination

17p. (p. 1-17)

Publisher

MDPI

ISSN

1999-4915

Rights Statement

The Author reserves all moral rights over the deposited text and must be credited if any re-use occurs. Documents deposited in OPAL are the Open Access versions of outputs published elsewhere. Changes resulting from the publishing process may therefore not be reflected in this document. The final published version may be obtained via the publisher’s DOI. Please note that additional copyright and access restrictions may apply to the published version.

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