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Vet J. 2012 Jul;193(1):67-72. doi: 10.1016/j.tvjl.2011.10.017. Epub 2011 Nov 23.

Rapid detection of foot-and-mouth disease virus using a field-portable nucleic acid extraction and real-time PCR amplification platform.

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1
Institute for Animal Health, Ash Road, Pirbright GU24 0NF, UK.

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.

PMID:
22115952
DOI:
10.1016/j.tvjl.2011.10.017
[Indexed for MEDLINE]
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