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マツイ ユウト
Yuuto Matsui
松井 優人 所属 川崎医科大学 医学部 基礎医学 微生物学 職種 助教 |
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| 論文種別 | 研究論文（学術雑誌） |
| 言語種別 | 英語 |
| 査読の有無 | 査読あり |
| 表題 | 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 |
| 掲載誌名 | 正式名：Journal of Virological Methods 略 称：J Virol Methods. |
| 掲載区分 | 国外 |
| 巻・号・頁 | 20(344) |
| 著者・共著者 | Kingkarn Boonsuya Seeyo, Kosuke Notsu, Nalinee Hongchumpon, Jeeranant Chottikamporn, Yuto Matsui, Janya Samanit, Alongkorn Pantumart, Parichat Ngansomsak, Mari Hirokawa, Kentaro Yamada, Lerdchai Chintapitaksakul, Naoaki Misawa |
| 発行年月 | 2026/05 |
| 概要 | Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals caused by foot-and-mouth disease virus (FMDV). FMDV can persist at low levels in probang fluid samples collected from clinically recovered cattle, posing challenges for sensitive detection and quantification. In this study, we developed and validated a one-step reverse transcription droplet digital PCR (RT-ddPCR) assay for the detection and absolute quantification of FMDV RNA. The analytical performance of RT-ddPCR was evaluated in comparison with the WOAH-recommended RT-qPCR assay. The RT-ddPCR assay demonstrated high analytical sensitivity, clear separation of positive and negative droplets, and strong quantitative agreement with RT-qPCR (Pearson's r = 0.997, p < 0.05). RT-ddPCR demonstrated analytical sensitivity equivalent to RT-qPCR, detecting FMDV RNA down to 1 × 100 TCID50/ml. Intra-assay repeatability showed low variability within runs, and inter-assay reproducibility across independent runs was comparable between RT-ddPCR and RT-qPCR. Application of the RT-ddPCR assay to probang fluid samples collected 6 and 10-11 months after FMD outbreaks revealed detectable levels of FMDV RNA in a subset of clinically recovered cattle. These findings indicate that one-step RT-ddPCR provides sensitive detection and absolute quantification of low-level FMDV RNA and represents a valuable complementary tool for investigating viral persistence and subclinical shedding in cattle. |