SPECIFIC DETECTION OF D-ISOLATES AND M-ISOLATES OF PLUM POX VIRUS BY IMMUNOENZYMATIC DETERMINATION OF PCR PRODUCTS

Molecular techniques based on the polymerase chain reaction (PCR) can provide rapid and sensitive diagnosis of plum pox virus (PPV), the cau sal agent of the devastating 'sharka' disease of stone fruit trees. Th e present study compared routine polymerase chain reaction (PCR) proce dures against a new system, PCR-ELISA (Boehringer Mannheim), which ena bles 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-sp ecific capture probes were also designed to identify the two major PPV isolates, D and M, without subsequent restriction fragment length pol ymorphism analysis of the PCR products. Optimisation of all parameters involved in the PCR-ELISA procedure are discussed and its advantages reported. (C) 1997 Elsevier Science B.V.