Specific detection of D- and M-isolates of plum pox virus by immunoenzymatic determination of PCR products

Abstract

Molecular techniques based on the polymerase chain reaction (PCR) can provide rapid and sensitive diagnosis of plum pox virus (PPV), the causal agent of the devastating `sharka' disease of stone fruit trees. The present study compared routine polymerase chain reaction (PCR) procedures against a new system, PCR-ELISA (Boehringer Mannheim), which enables immunoenzymatic detection of PCR products. The results show that this hybridisation system ensures fast and more sensitive detection of PPV associated with stone fruit trees and herbaceous hosts. Strain-specific capture probes were also designed to identify the two major PPV isolates, D and M, without subsequent restriction fragment length polymorphism analysis of the PCR products. Optimisation of all parameters involved in the PCR-ELISA procedure are discussed and its advantages reported.