Adaptation of a <i>Phytophthora ramorum</i> Real-Time Polymerase Chain Reaction Assay Based on a Mitochondrial Gene Region for Use on the Cepheid SmartCycler

Fig. 1. Sensitivity of the mitochondrial gene region-based Phytophthora ramorum detection assay using the Cepheid SmartCycler. (A) Real-time PCR amplification profile for a representative dilution series of DNA extracted from Phytophthora ramorum isolate 288. (B) Standard curve of cycle threshold (Ct) values calculated from serial dilutions of P. ramorum DNA tested with the P. ramorum primer and probe set. (C) Standard curve of Ct values calculated from serial dilutions of P. ramorum DNA spiked with DNA extracted from a healthy azalea and tested with the P. ramorum primer and probe set. (D) Standard curve of Ct values calculated from serial dilutions of P. ramorum DNA spiked with DNA extracted from a healthy azalea and tested with both the P. ramorum and plant primer and probe sets. Three separate DNA dilutions were produced for each experiment and duplicate PCR reactions were tested with each dilution (n = 6). Sigma Plot version 10.0 (Systat Software Inc., Point Richmond, CA) was used for statistical analyses; standard error bars are indicated.



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Adaptation of a Phytophthora ramorum Real-Time Polymerase Chain Reaction Assay Based on a Mitochondrial Gene Region for Use on the Cepheid SmartCycler