ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Veterinary Infectious Diseases
Development of a novel p72 gene-based loop-mediated isothermal amplification assay for the rapid detection of African swine fever virus in animal feed
Provisionally accepted
Beilei Ge1*
Kelly J Domesle1
Janine A Simmons2
Shenia Young1David A Brake2
Ryan McDonald1Haile F Yancy1Lindsay R Gabbert2
John Gerard Neilan1
Chris Whitehouse1- 1US Food and Drug Administration Center for Veterinary Medicine, Rockville, United States
- 2US Department of Homeland Security Science and Technology Directorate, Washington, United States
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Rapid and reliable detection of African swine fever virus (ASFV), the causative agent of often fatal African swine fever, in animal feed is critical for implementing timely emergency control measures. We developed a novel loop-mediated isothermal amplification (LAMP) assay targeting the ASFV p72 gene (B646L, encoding the major capsid protein) compatible with animal feed. Assay performance (referred to as LAMP1 in this study) was evaluated in comparison with a previously published topoisomerase II gene-based LAMP assay (LAMP2) and three p72-based real-time PCR assays (two recommended by the World Organisation for Animal Health [WOAH] and the third one developed and currently used by the U.S. Department of Agriculture). LAMP1 was the fastest with positive results obtained in as early as 3.8 min (compared to at least 5.5 min for LAMP2 and 20 cycles for real-time PCRs [~ 15 min]). These assays could detect ASFV from 10-1 to 102 copies using synthetic DNA. LAMP1 detected as low as 101 TCID50/ml of ASFV BA71V stock and had comparable performance to the USDA real-time PCR assay using both inclusivity (36 ASFV synthetic DNAs and isolates) and exclusivity (13 porcine viruses) panels. Applying six different DNA extraction methods to a variety of animal feed sample types (e.g., complete swine feed, soybean meal), variable yet mostly limited assay inhibitions were observed. When swine feed was inoculated with the ASFV BA71V stock at 105.1 TCID50/g, the newly developed LAMP1 assay reliably detected the virus within 7 min in contrast to at least 20 min by the USDA real-time PCR (29 cycles). Further validation of this novel p72-based LAMP assay in animal feed will pave the way for its adoption as a rapid screening tool in ASFV feed surveillance, as well as potential application in outbreak response and recovery efforts, to safeguard the nation's animal feed supply.
Keywords: African Swine Fever Virus, Animal Feed, detection, loop-mediated isothermalamplification, major capsid protein
Received: 06 Aug 2025; Accepted: 03 Nov 2025.
Copyright: © 2025 Ge, Domesle, Simmons, Young, Brake, McDonald, Yancy, Gabbert, Neilan and Whitehouse. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Beilei Ge, beilei.ge@fda.hhs.gov
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