Immunoenzymatic Detection of PCR Products for the Identification of Phytoplasmas in Plants

Polymerase chain reaction (PCR, with universal and specific primers designed on rRNA genes) provides a rapid, reliable method of diagnosing phytoplasmas (formerly mycoplasma-like organisms) in plants. However, to attain a better identification of these prokaryotes, it is often necessary to digest the PCR products with restriction endonucleases or to hybridize them with specific probes. The present study compared routine procedures for detecting PCR products against a new system, PCRELISA (Boehringer Mannheim), which enables immunoenzymatic detection of PCR products. The results show that this new system provides fast and highly sensitive detection of several phytoplasmas associated with certain trees and shrubs. Optimization of all parameters involved in the PCR-ELISA procedure and its advantages are reported and discussed