title: Rapid detection of foot-and-mouth disease virus using a field-portable nucleic acid extraction and real-time PCR amplification platform.
abstract: Rapid and accurate field diagnostic tools can be used to support clinical diagnosis during outbreaks of exotic livestock diseases. This study evaluated a mobile PCR amplification platform that performs RNA extraction, real-time reverse-transcription PCR (rRT-PCR) and interpretation of results without operator intervention. Initial studies showed that there was equivalence between the detection limit generated by RNA extracted using the mobile platform and an automated laboratory-based system. In subsequent studies, two validated laboratory-based foot-and-mouth disease virus (FMDV)-specific rRT-PCRs were transferred onto the mobile platform and all assay steps (RT incubation and PCR amplification) were performed with non-lyophilised reagents using an optimised protocol of less than 60 min. The limit of detection of the rRT-PCR on the mobile PCR platform was equivalent to an automated laboratory-based assay used for routine diagnosis of FMDV and there was concordance between these methods for results generated using samples in a reference laboratory proficiency panel. Future studies will be directed at the development and validation of commercially-viable consumables using lyophilised PCR reagents for FMDV and the evaluation of this technology in FMD endemic countries using field samples.
authors: Institute for Animal Health, Ash Road, Pirbright GU24 0NF, UK.
authors: Mikidache
authors: M
authors: Madi
authors: Andrea
authors: A
authors: Hamilton
authors: David
authors: D
authors: Squirrell
authors: Valerie
authors: V
authors: Mioulet
authors: Phillip
authors: P
authors: Evans
authors: Martin
authors: M
authors: Lee
authors: Donald P
authors: DP
authors: King
mesh_terms: D000818:Animals; D057205:Automation, Laboratory / Q000379:methods; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology; D005536:Foot-and-Mouth Disease / Q000175:diagnosis*; D029544:Foot-and-Mouth Disease Virus / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification; D011933:Reagent Kits, Diagnostic / Q000662:veterinary; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*
chemical_list: D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic