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United States Department of Agriculture

Agricultural Research Service

Research Project: CHARACTERIZATION & EPIDEMIOLOGY OF CITRUS TRISTEZA VIRUS & OTHER INVASIVE & EMERGING GRAFT-TRANSMISSIBLE DISEASES OF CITRUS IN CALIFORNIA Title: Development of a High Throughput Assay for Rapid and Accurate 10-Plex Detection of Citrus Pathogens

Authors
item Wang, Jinbo -
item Vidalakis, Georgios -
item Lee, Richard -
item Yokomi, Raymond

Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: May 14, 2012
Publication Date: July 1, 2012
Citation: Wang, J., Vidalakis, G., Lee, R., Yokomi, R.K. 2012. Development of a High Throughput Assay for Rapid and Accurate 10-Plex Detection of Citrus Pathogens. Phytopathology. 102:S4.131.

Technical Abstract: The need to reliably detect and identify multiple plant pathogens simultaneously, especially in woody perennial hosts, has led to development of new molecular diagnostic approaches. In this study, a Luminex-based system was developed that provided a robust and sensitive test for simultaneous detection, identification and quantification of nine citrus pathogen targets. The assay included a target for a citrus housekeeping gene, Nad5, as an internal control. The protocol developed here is the first to successfully detect up to 10 targets in one multiplex assay. A procedure for high throughput robotic extraction and purification of pathogen nucleic acid targets, optimized for citrus tissue, was developed and used with the Luminex-based system to increase uniformity and cost effectiveness of the test. The assay is user friendly and can be applied to detect endemic and/or exotic plant pathogens in quarantine or certification programs.

Last Modified: 11/25/2014
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