Abstract
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.
Highlights
African swine fever (ASF) is a highly contagious, viral disease in swine characterised by fever, haemorrhages and a high mortality rate [1]
The African swine fever virus (ASFV) Loop-mediated isothermal amplification (LAMP) assay was successfully optimized for both primer concentration ratio and temperature
The ASFV LAMP assay was successfully used to detect the presence of ASFV in multiple serum samples from Baucau, Timor-Leste
Summary
African swine fever (ASF) is a highly contagious, viral disease in swine characterised by fever, haemorrhages and a high mortality rate [1]. The disease is caused by African swine fever virus (ASFV), the only member of the genus Asfivirus family Asfarviridae [1]. The transboundary nature of the disease saw outbreaks from Africa into Eastern Europe in 2007 (genotype II, Georgia 2007) [2] and has since spread throughout the world. The continued movement of the virus saw large outbreaks in China in August 2018 [3] and other Asian countries such as Vietnam [4], Cambodia, the Philippines [5] and Korea [6] in 2019. Once established in a region, the virus can rapidly spread throughout a swine population. Death can occur in 2–10 days, with mortality rates
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
