Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the Rapid and Sensitive Detection of Yam mosaic virus

• Published in: Viruses Vol. 15; no. 7; p. 1592
• Main Authors: Festus, Ruth O, Seal, Susan E, Prempeh, Ruth, Quain, Marian D, Silva, Gonçalo
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 21.07.2023; MDPI
• Subjects: Food security; Fourier transforms; Gene amplification; Genomes; isothermal amplification; Laboratories; Phylogenetics; Polyethylene glycol; Polymerase chain reaction; Primers; Propagation; Reverse transcription; RT-LAMP; seed systems; Seeds; Thermal cycling; virus diagnostics; Viruses; yam; Yam mosaic virus; New Zealand; United Kingdom > UK; United States > US; West Africa; RT-LAMP; seed systems; Yam mosaic virus; yam; West Africa; isothermal amplification; virus diagnostics
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v15071592

Yam ( 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 (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.