Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the rapid and sensitive detection of Yam mosaic virus
Festus, Ruth, Seal, Susan ORCID: https://orcid.org/0000-0002-3952-1562, Prempeh, Ruth, Quain, Marian D. and Silva, Goncalo ORCID: https://orcid.org/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:10.3390/v15071592)
Preview |
PDF (Publisher VoR)
43442_SILVA_Improved_Reverse_Transcription_Loop_Mediated_Isothermal_Amplification_RT-LAMP.pdf - Published Version Available under License Creative Commons Attribution. Download (3MB) | Preview |
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.
Actions (login required)
View Item |
Downloads
Downloads per month over past year