|WANG, JINBO - University Of California|
|VIDALAKIS, GEORGIOS - University Of California|
|LEE, RICHARD - University Of Florida|
|Yokomi, Raymond - Ray|
Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: 5/14/2012
Publication Date: 7/1/2012
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.