COMPARISON OF REAL-TIME PCR PROTOCOLS IN DETECTION AND QUANTIFICATION OF FRUIT TREE 16SRX GROUP PHYTOPLASMAS

In this work, two real-time PCR protocols based on intercalating dye and two on hydrolysis probes were tested using field collected fruit tree samples infected by 16SrX group (AP, PD and ESFY) phytoplasmas. Specificity and sensitivity of protocols and amplification efficiency were the main testing parameters. Results of real-time PCR protocols were compared to nested PCR. All real-time PCR protocols confirmed their specificity of detection. All real-time PCR protocols were 10-100 times more sensitive than nested PCR. Afterall real-time PCR protocols based on hydrolysis probes were 10 times more sensitive than protocols based on intercalating dyes. Among protocols based on hydrolysis probes, slightly better detection characteristics were shown by protocol by CHRISTENSEN et al. (2004).