Development of a real-time RT-PCR system applicable for rapid and pen-side diagnosis of foot-and-mouth disease using a portable device, PicoGene® PCR1100

• Published in: Journal of virological methods Vol. 319; p. 114753
• Main Authors: Matsui, Yuto, Chottikamporn, Jeeranant, Ungvanijban, Sahawatchara, Seeyo, Kingkarn Boonsuya, Vitoonpong, Ratchaneekorn, Suwankitwat, Nutthakarn, Songkasupa, Tapanut, Norimine, Junzo, Yamada, Kentaro, Chintapitaksakul, Lerdchai, Misawa, Naoaki
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.09.2023
• Subjects: Animals; Foot-and-mouth disease; Foot-and-Mouth Disease - diagnosis; Foot-and-Mouth Disease Virus - genetics; Pen-side diagnosis; Portable real-time PCR device; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral - genetics; Sensitivity and Specificity; PBS; Pen-side diagnosis; RRLFMD; Portable real-time PCR device; LOD50; rRT-PCR; RT-LAMP; WOAH; FMDV; RT-iiPCR; RT-RPA; Foot-and-mouth disease; FMD
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2023.114753

Foot-and-mouth disease (FMD) is a highly contagious viral vesicular disease, causing devastating losses to the livestock industry. A diagnostic method that enables quick decisions is required to control the disease, especially in FMD-free countries. Although conventional real-time reverse transcription polymerase chain reaction (RT-PCR) is a highly sensitive method widely used for the diagnosis of FMD, a time lag caused by the transport of samples to a laboratory may allow the spread of FMD. Here, we evaluated a real-time RT-PCR system using a portable PicoGene PCR1100 device for FMD diagnosis. This system could detect the synthetic FMD viral RNA within 20 min with high sensitivity compared to a conventional real-time RT-PCR. Furthermore, the Lysis Buffer S for crude nucleic extraction improved the viral RNA detection of this system in a homogenate of vesicular epithelium samples collected from FMD virus-infected animals. Furthermore, this system could detect the viral RNA in crude extracts prepared using the Lysis Buffer S from the vesicular epithelium samples homogenized using a Finger Masher tube, which allows easy homogenization without any equipment, with a high correlation compared to the standard method. Thus, the PicoGene device system can be utilized for the rapid and pen-side diagnosis of FMD. •A portable real-time PCR device was evaluated for foot-and-mouth disease (FMD) diagnosis.•A rapid diagnosis system that can be used in the field has been developed.•This system showed a high sensitivity comparable to the gold standard method.