Testing for Sudden Oak Death – PCR Method

DNA Extraction
The polymerase chain reaction (PCR) method is used to detect the presence of P. ramorum in plant material. Scientific paper. Infected plant material is placed in a sample tube with a small glass bead, freeze dried, and pulverized in a tissue homogenizer.
The plant material is extensively processed to extract, isolate, and purify the target DNA. The DNA containing solutions are transfered to a microtiter plate to increase efficiency and assay throughput.
PCR Setup
Real Time PCR Machine
The thermocycler (Real-Time PCR machine) is used with P. ramorum-specific primers to amplify and simultaneously quantify regions of the DNA that are unique to P. ramorum. Scientific articles: Hayden 2006, Chandelier 2006, Martin 2009.


Pros and Cons of the PCR method:
Pro:
– The gold standard for P. ramorum detection.
– Highly specific for species identification.
– Eliminates false negative and false positive results.

Con:
– Expensive compared to culturing and EIA methods.
– Requires specialized equipment and training.
– Relatively slow compared to EIA methods.

Jump to Other SOD Diagnostic Methods:
Immunostrips – Culturing – EIA Analysis – PCR Analysis

The UC Berkeley Forest Pathology and Mycology Laboratory does not provide a SOD testing service. For information on SOD testing please visit www.suddenoakdeath.org

This activity possible thanks to funding from:
USDA Forest Service, State and Private Forestry
The Gordon and Betty Moore Foundation
Agdia, Inc. http://www.agdia.com

Disclaimer: Mention of any company, trade name, or commercial product does not constitute endorsement by the University of California or recommendation for use.