DEVELOPING A METHOD TO DETECT XYLELLA FASTIDIOSA IN THE GLASSY-WINGED SHARPSHOOTER

• Author(s): Bextine, Blake; Blua, Matthew; Redak, Richard;
• Abstract: A rapid and reproducible technique to detect Xylella fastidiosa (Xf) in the glassy-winged sharpshooter (GWSS) is important for epidemiological studies, and monitoring programs in support of Pierces disease management. Such a technique must be amenable to large samples sizes, while remaining sensitive enough to detect pathogen DNA in low amounts. In this study we have improved the speed of tissue extraction by developing a simple vacuum step that replaces labor and time-intensive tissue maceration, and is compatible with manufactured DNA extraction kits and a SYBR Green based real-time (QRT) PCR system. No statistical differences in the ability to detect Xf were found among samples that were extracted using traditional maceration vs. our vacuum extraction method. Further experiments using our vacuum extraction methods detected no significant differences among samples immediately extracted, or stored for 10 d at -4C, dry or in mineral oil. In another experiment we placed Xf -fed GWSS on yellow sticky cards in a sunny location for 0 to 6 d. We found that there was no significant reduction in our detection capabilities for insects left on the cards.
• Publication Date: Dec 2004
• Journal: 2004 Pierce's Disease Research Symposium