Please use this identifier to cite or link to this item: http://hdl.handle.net/1893/18312
Appears in Collections:Aquaculture Journal Articles
Peer Review Status: Refereed
Title: Recombinase polymerase amplification assay for rapid detection of Rift Valley fever virus
Authors: Euler, Milena
Wang, Yongjie
Nentwich, Oliver
Piepenburg, Olaf
Hufert, Frank T
Weidmann, Manfred
Contact Email: m.w.weidmann@stir.ac.uk
Keywords: RPA
Recombinase polymerase amplification
Rift Valley fever virus
Issue Date: Aug-2012
Publisher: Elsevier
Citation: Euler M, Wang Y, Nentwich O, Piepenburg O, Hufert FT & Weidmann M (2012) Recombinase polymerase amplification assay for rapid detection of Rift Valley fever virus, Journal of Clinical Virology, 54 (4), pp. 308-312.
Abstract: Background: Detection of nucleic acids of Rift Valley fever virus (RVFV) has been shown to be useful in field diagnostics. Objectives: To develop an isothermal ‘recombinase polymerase amplification (RPA)' assay on an ESEquant tubescanner device. Study design: RPA was adapted for RNA amplification by first developing a two-step and then a one-step-RT-RPA protocol. Several RT enzymes were tested and the best sensitivity was achieved using Transcriptor (Roche). Finally an RT-RPA pellet containing a recombinant MuLV was tested in RVFV one-step-RT-RPA. Results: The one-step-RT-RPA assay showed a sensitivity of 19 molecules detected as determined by probit analysis of eight runs using a RVFV S-segment based quantitative RNA standard and detected 20 different RVFV strains. The assays showed no cross detection of the human genome and several agents of a typical biothreat panel. It performed almost as good as the assay using glycerol buffer based Transcriptor albeit at a cost of 1-log10 step in sensitivity. The presented combination of one-step-RT-RPA and portable fluorescence reading device could be a useful tool for field or point of care diagnostics.
Type: Journal Article
URI: http://hdl.handle.net/1893/18312
DOI Link: http://dx.doi.org/10.1016/j.jcv.2012.05.006
Rights: The publisher does not allow this work to be made publicly available in this Repository. Please use the Request a Copy feature at the foot of the Repository record to request a copy directly from the author. You can only request a copy if you wish to use this work for your own research or private study.
Affiliation: University of Gottingen, Georg-August University
University of Gottingen, Georg-August University
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University of Gottingen, Georg-August University
Aquaculture

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