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A loop-mediated isothermal amplification (lamp) assay for rapid identification of Bemisia tabaci

Blaser, Simon and Diem, Hanspeter and von Felten, Andreas and Gueuning, Morgan and Andreou, Michael and Boonham, Neil and Tomlinson, Jennifer and Müller, Pie and Utzinger, Jürg and Frey, Beatrice and Frey, Jürg E. and Bühlmann, Andreas. (2018) A loop-mediated isothermal amplification (lamp) assay for rapid identification of Bemisia tabaci. Journal of visualized experiments (140). e58502.

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Abstract

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many countries around the world. Severe yield losses are caused by direct feeding, and even more importantly, also by the transmission of more than 100 harmful plant pathogenic viruses. As for other invasive pests, increased international trade facilitates the dispersal of B. tabaci to areas beyond its native range. Inspections of plant import products at points of entry such as seaports and airports are, therefore, seen as an important prevention measure. However, this last line of defense against pest invasions is only effective if rapid identification methods for suspicious insect specimens are readily available. Because the morphological differentiation between the regulated B. tabaci and close relatives without quarantine status is difficult for non-taxonomists, a rapid molecular identification assay based on the loop-mediated isothermal amplification (LAMP) technology has been developed. This publication reports the detailed protocol of the novel assay describing rapid DNA extraction, set-up of the LAMP reaction, as well as interpretation of its read-out, which allows identifying B. tabaci specimens within one hour. Compared to existing protocols for the detection of specific B. tabaci biotypes, the developed method targets the whole B. tabaci species complex in one assay. Moreover the assay is designed to be applied on-site by plant health inspectors with minimal laboratory training directly at points of entry. Thorough validation performed under laboratory and on-site conditions demonstrates that the reported LAMP assay is a rapid and reliable identification tool, improving the management of B. tabaci.
Faculties and Departments:09 Associated Institutions > Swiss Tropical and Public Health Institute (Swiss TPH)
09 Associated Institutions > Swiss Tropical and Public Health Institute (Swiss TPH) > Department of Epidemiology and Public Health (EPH) > Eco System Health Sciences > Health Impact Assessment (Utzinger)
09 Associated Institutions > Swiss Tropical and Public Health Institute (Swiss TPH) > Department of Epidemiology and Public Health (EPH) > Health Interventions > Vector Control Centre (Müller)
UniBasel Contributors:Müller, Pie and Utzinger, Jürg and Blaser, Simon
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:JoVE
ISSN:1940-087X
Note:Publication type according to Uni Basel Research Database: Journal article
Language:English
Identification Number:
Last Modified:26 Feb 2019 08:19
Deposited On:26 Feb 2019 08:19

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