Difficulties with conventional phytoplasma diagnostic using PCR/RFLP analyses

Polymerase chain reaction (PCR) with subsequent restriction fragment length polymorphism (RFLP) analysis is often used for phytoplasma identification and classification. Although these techniques are very sensitive and specific, in some cases, nonspecific reactions, false positives and negatives results, as well as unusual or illegible profiles after RFLP analyses, amplification of plant host ́s DNA or other difficulties occurred. Experiences with suitability of positive and negative controls integration in PCR, evaluation of critical samples and other difficulties in phytoplasma PCR/RFLP identification are reported.