SIMULTANEOUS DETECTION AND TYPING OF PLUM POX POTYVIRUS (PPV) ISOLATES BY HEMINESTED-PCR AND PCR-ELISA

Two techniques for simultaneous detection and typing of plum pox potyv irus (PPV) isolates belonging to the D or M serotypes, heminested PCR (H-PCR) and PCR-ELISA, have been developed. Ten PPV isolates typed usi ng PPV-D and PPV-M specific monoclonal antibodies by ELISA-DASI were u sed to validate these two methods. The results obtained show a complet e coincidence of the nucleic acid-based techniques with the serologica l data. When serial dilutions of infected plant extracts were assayed, H-PCR and PCR-ELISA were found to be 100 rimes more sensitive than th e more conventional immunocapture-PCR (IC-PCR) assay. Testing of 228 P PV-infected fruit tree samples coming from different hosts and locatio ns indicated that so fat-only PPV type D appears to be present in Spai n and in Chile. Coupled with print-capture sample preparation (Olmos e t al., Nucl. Acids Res. 24, 2192-2193, 1996) the increased sensitivity provided by heminested-PCR allowed the detection of PPV targets of D and M types, in wingless individuals of the aphid vector Aphis gossypi i. (C) 1997 Elsevier Science Ireland Ltd.