International Collaborative Research Achievement: Development of a Highly Sensitive Molecular
Diagnostic Method for Foot-and-Mouth Disease Virus
― Published in the Journal of Virological Methods ―
The Center for Animal Disease Control (CADIC), University of Miyazaki, in collaboration with the Regional Reference Laboratory for Foot-and-Mouth Disease, Department of Livestock Development, Thailand, has developed a novel molecular diagnostic method for highly sensitive detection and absolute quantification of foot-and-mouth disease virus (FMDV) RNA using a one-step reverse transcription droplet digital PCR (RT-ddPCR) assay. The research findings have been published in the international scientific journal Journal of Virological Methods.
Foot-and-mouth disease (FMD) is one of the most important transboundary animal diseases affecting cloven-hoofed animals such as cattle, pigs, sheep, and goats. The disease causes severe economic losses to the livestock industry and international trade. Because FMD remains endemic in many Asian countries, the development of rapid, highly sensitive diagnostic technologies is critical for disease surveillance and control.
In this study, the research team developed a novel droplet digital PCR (ddPCR)-based diagnostic assay capable of detecting extremely low levels of FMDV RNA. The assay demonstrated analytical sensitivity comparable to the WOAH-recommended real-time RT-PCR (RT-qPCR) method while enabling absolute quantification of viral RNA without the need for standard curves. The method also showed high reproducibility and robustness for low-copy-number samples.
Using probang fluid (oropharyngeal fluid) samples collected from naturally infected cattle in Thailand, the study demonstrated that very low levels of FMDV RNA could still be detected up to 10-11 months after field outbreaks. These findings suggest that the RT-ddPCR assay may serve as a valuable tool for monitoring persistent low-level viral RNA in clinically recovered cattle.
The study is expected to contribute not only to improved FMD surveillance and post-outbreak monitoring but also to strengthening transboundary animal disease control strategies in the ASEAN region.
This research was conducted as part of the JST-JICA SATREPS project: "Acceleration of Livestock Revolution in ASEAN for a Kitchen of the World: Development of Novel Technologies Yielding Stable Livestock Production and Food Safety".
Collecting probang fluid
Publication Information
Kingkarn Boonsuya Seeyo, Kosuke Notsu, Nalinee Hongchumpon, Jeeranant Chottikamporn, Yuto Matsui, Janya Samanit, Alongkorn Pantumart, Parichat Ngansomsak, Mari Hirokawa, Kentaro Yamada, Lerdchai Chintapitaksakul, and Naoaki Misawa "Development and validation of a one-step RT-ddPCR assay for sensitive and absolute quantification of foot-and-mouth disease virus RNA in probang fluid" Journal of Virological Methods, Volume 344 (2026)
DOI: https://doi.org/10.1016/j.jviromet.2026.115411
