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Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification

Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification

Silva, Gonçalo, Bömer, Moritz, Nkere, Chukwuemeka, Kumar, P. Lava and Seal, Susan E. (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: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 / Department / 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:
Last Modified: 11 Apr 2017 13:53
Selected for GREAT 2016: GREAT a
Selected for GREAT 2017: None
Selected for GREAT 2018: None
URI: http://gala.gre.ac.uk/id/eprint/13756

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