Silva, Gonçalo ORCID: 0000-0001-5544-2947 , Bömer, Moritz ORCID: 0000-0003-1003-9622 , Nkere, Chukwuemeka, Kumar, P. Lava and Seal, Susan E. ORCID: 0000-0002-3952-1562 (2015) Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification. Journal of Virological Methods, 222. pp. 138-144. ISSN 0166-0934 (doi:https://doi.org/10.1016/j.jviromet.2015.06.011)

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Abstract

Yam mosaic virus (YMV; genus Potyvirus) is considered to cause the most economically important viral disease of yams (Dioscorea spp.) in West Africa which is the dominant region for yam production globally.

Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps.

In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase amplification (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/μl of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37 °C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratories.

Item Type:	Article
Additional Information:	[1] Acknowledgements (funding): The authors acknowledge the support of this work by the Bill & Melinda Gates Foundation under the ‘Development of On-farm Robust Diagnostic Toolkits for Yam Viruses’ grant to NRI and the ‘Yam Improvement for Income and Food Security in West Africa (YIIFSWA)’ grant to IITA.
Uncontrolled Keywords:	yam, dioscorea spp., YMV, recombinase polymerase amplification, diagnosis
Subjects:	Q Science > Q Science (General) Q Science > QR Microbiology
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
Related URLs:	Publisher
Last Modified:	30 Apr 2020 16:34
URI:	http://gala.gre.ac.uk/id/eprint/13756

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