Silva, Gonçalo ORCID: 0000-0001-5544-2947, Oyekanmi, Joshua, Nkere, Chukwuemeka K., Bömer, Moritz ORCID: 0000-0003-1003-9622, Kumar, Lava P. and Seal, Susan ORCID: 0000-0002-3952-1562 (2018) Rapid detection of potyviruses from crude plant extracts. Analytical Biochemistry, 546. pp. 17-22. ISSN 0003-2697 (Print), 1096-0309 (Online) (doi:https://doi.org/10.1016/j.ab.2018.01.019)

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Abstract

Potyviruses (genus Potyvirus; family Potyviridae) are widely distributed and represent one of the most economically important genera of plant viruses. Therefore, their accurate detection is a key factor in developing efficient control strategies. However, this can sometimes be problematic particularly in plant species containing high amounts of polysaccharides and polyphenols such as yam (Dioscorea spp.). Here, we report the development of a reliable, rapid and cost-effective detection method for the two most important potyviruses infecting yam based on reverse transcription-recombinase polymerase amplification (RT-RPA). The developed method, named ‘Direct RT-RPA’, detects each target virus directly from plant leaf extracts prepared with a simple and inexpensive extraction method avoiding laborious extraction of high-quality RNA. Direct RT-RPA enables the detection of virus-positive samples in under 30 min at a single low operation temperature (37 °C) without the need for any expensive instrumentation. The Direct RT-RPA tests constitute robust, accurate, sensitive and quick methods for detection of potyviruses from recalcitrant plant species. The minimal sample preparation requirements and the possibility of storing RPA reagents without cold chain storage, allow Direct RT-RPA to be adopted in minimally equipped laboratories and with potential use in plant clinic laboratories and seed certification facilities worldwide.

Item Type:	Article
Additional Information:	Special Issue of Analytical Biochemistry entitled “Recombinase Polymerase Amplification: Applications and Advances" *** © 2018 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/).
Uncontrolled Keywords:	Yam; Recombinase polymerase amplification; Potyvirus; Isothermal amplification; Rapid diagnostics
Subjects:	S Agriculture > S Agriculture (General)
Faculty / School / Research Centre / Research Group:	Faculty of Engineering & Science Faculty of Engineering & Science > Natural Resources Institute Faculty of Engineering & Science > Natural Resources Institute > Agriculture, Health & Environment Department
Last Modified:	29 Apr 2020 14:02
URI:	http://gala.gre.ac.uk/id/eprint/19187

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