Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses.

Autor: Wanjala BW; International Potato Center, SSA Regional Office, PO Box 25171, 00603, Nairobi, Kenya; Jomo Kenyatta University of Agriculture and Technology, P.O. Box 62000, 00200, Nairobi, Kenya. Electronic address: bramwel.wanjala@kalro.org., Ateka EM; Jomo Kenyatta University of Agriculture and Technology, P.O. Box 62000, 00200, Nairobi, Kenya. Electronic address: eateka@agr.jkuat.ac.ke., Miano DW; University of Nairobi, P.O. Box: 30197, 00100, Nairobi, Kenya. Electronic address: dmiano@uonbi.ac.ke., Fuentes S; International Potato Center, Avenida La Molina 1895, La Molina, Apartado Postal 1558, Lima, Peru. Electronic address: s.fuentes@cgiar.org., Perez A; International Potato Center, Avenida La Molina 1895, La Molina, Apartado Postal 1558, Lima, Peru. Electronic address: a.perez@cgiar.org., Low JW; International Potato Center, SSA Regional Office, PO Box 25171, 00603, Nairobi, Kenya. Electronic address: j.low@cgiar.org., Kreuze JF; International Potato Center, Avenida La Molina 1895, La Molina, Apartado Postal 1558, Lima, Peru. Electronic address: j.kreuze@cgiar.org.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2021 Dec; Vol. 298, pp. 114301. Date of Electronic Publication: 2021 Sep 21.
DOI: 10.1016/j.jviromet.2021.114301
Abstrakt: Globally, Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) occur frequently and in combination cause sweetpotato virus disease (SPVD). Many viral diseases are economically important and negatively impact the production and movement of germplasm across regions. Rapid detection of viruses is critical for effective control. Detection and quantification of viruses directly from sweetpotato remains a challenge. Current diagnostic tests are not sensitive enough to reliably detect viruses directly from the plant or require expensive laboratory equipment and expertise to perform. We developed a simple and rapid loop-mediated isothermal amplification (LAMP) assay for the detection of SPFMV, SPCSV and begomoviruses related to sweet potato leaf curl virus (SPLCV). Laboratory validation recorded 100 % diagnostic sensitivity for all the three viruses. The LAMP assays were customized for field testing using a lyophilized thermostable isothermal master mix in a ready-to-use form that required no cold chain. The average time to positivity (TTP) was: SPFMV 5-30 min, SPCSV 15-43 min s and begomoviruses 28-45 mins. LAMP on-site testing results were comparable to PCR and RT-PCR confirmatory laboratory tests. The LAMP assay is a powerful tool for rapid sweetpotato virus detection at a reasonable cost and thus could serve as quality control systems for planting materials.
(Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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