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Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the rapid and sensitive detection of Yam mosaic virus

Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the rapid and sensitive detection of Yam mosaic virus

Festus, Ruth, Seal, Susan ORCID: 0000-0002-3952-1562 , Prempeh, Ruth, Quain, Marian D. and Silva, Goncalo ORCID: 0000-0001-5544-2947 (2023) Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the rapid and sensitive detection of Yam mosaic virus. Viruses, 15 (7):1592. pp. 1-17. ISSN 1999-4915 (Online) (doi:https://doi.org/10.3390/v15071592)

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Abstract

Yam (Dioscorea spp.) productivity is constrained significantly by the lack of a formal seed system. Vegetative propagation, through tuber setts as ‘seed’ yams, encourages the recycling of virus-infected planting materials, contributing to high virus incidence and yield losses. Efforts are ongoing to increase the production of high-quality seed yams in a formal seed system to reduce virus-induced yield losses and enhance the crop’s productivity and food security. Specific and sensitive diagnostic tests are imperative to prevent the multiplication of virus-infected materials contributing to a sustainable seed yam certification system. During routine indexing of yam accessions, discrepancies were observed between the results obtained from the reverse transcription loop-mediated isothermal amplification (RT-LAMP) test and those from reverse transcription polymerase chain reaction (RT-PCR); RT-LAMP failed to detect Yam mosaic virus (YMV) in some samples that tested positive by RT-PCR. This prompted the design of a new set of LAMP primers, YMV1-OPT primers. These primers detected as little as 0.1 fg/µL of purified RNA obtained from a YMV-infected plant, a sensitivity equivalent to that obtained with RT-PCR. RT-LAMP using YMV1-OPT primers is recommended for all future virus-indexing of seed yams for YMV, offering a rapid, sensitive, and cost-effective approach.

Item Type: Article
Additional Information: This article belongs to the Section Viruses of Plants, Fungi and Protozoa.
Uncontrolled Keywords: yam; seed systems; Yam mosaic virus; virus diagnostics; isothermal amplification; RT-LAMP; West Africa
Subjects: Q Science > Q Science (General)
Q Science > QR Microbiology > QR355 Virology
S Agriculture > SB Plant culture
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
Faculty of Engineering & Science > Natural Resources Institute > Molecular Virology and Entomology Research Group
Last Modified: 25 Jul 2023 09:06
URI: http://gala.gre.ac.uk/id/eprint/43442

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