1. Rapid detection of foot-and-mouth disease virus using a field-portable nucleic acid extraction and real-time PCR amplification platform.
- Author
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Madi, Mikidache, Hamilton, Andrea, Squirrell, David, Mioulet, Valerie, Evans, Phillip, Lee, Martin, and King, Donald P.
- Subjects
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LIVESTOCK diseases , *FOOT & mouth disease , *REVERSE transcriptase polymerase chain reaction , *BIOLOGICAL reagents , *POLYMERASE chain reaction - 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 rain. 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. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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