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Comparison of Monoclonal Antibodies and Polymerase Chain Reaction Assays for the Typing of Isolates Belonging to the D and M Serotypes of Plum Pox Potyvirus

First, fourth, seventh, eighth, and twelfth authors: Station de Pathologie Végétale, INRA, BP81, 33883 Villenave d'Ornon Cedex, France; second, fifth, sixth, and ninth authors: Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Apartado oficial, 46113 Moncada, Valencia, Spain; third and eleventh authors: Pathologie Végétale, INRA, 2 Place Viala, 34060 Montpellier Cedex 01, France; and tenth author: Dipartimento di Protezione delle Piante, Universita degli Studi and Centro di Studio del CNR sui Virus e Virosi delle Colture Mediteranee, Via Amendola 165/A, 70126 Bari, Italy

Plum pox potyvirus (PPV) isolates may be divided into four groups separated by serological, molecular, and epidemiological differences. Monoclonal antibodies specific for the two major groups of isolates, represented by the D and M serotypes of the virus, have been obtained. Polymerase chain reaction (PCR)-based assays allowing the direct detection and differentiation of PPV isolates have also been developed. We now report on a large-scale comparison of these two typing approaches. The results obtained show an overall excellent correlation between the results obtained in indirect double-antibody sandwich enzyme-linked immunosorbent assay using PPV-D— and PPV-M—specific monoclonal antibodies and those derived from either specific PCR assays or restriction fragment length polymorphism analysis of PCR fragments. Without exception, all isolates reacting positively with the PPV-M—specific monoclonal antibody were found to belong to the M serotype using the PCR-based assays, while 51 out of 53 isolates recognized by the D-specific monoclonal antibodies belonged to the D serotype according to the PCR typing results. However, failure to react with a specific monoclonal antibody did not prove as effective a predictor of the serotype of the isolate analyzed. In a few cases, the results obtained with the various techniques diverged, indicating low level variability of the epitopes recognized by the serotype-specific monoclonal antibodies. Isolates belonging to the two minor groups of PPV (El Amar and Cherry) also gave divergent results, indicating that the current typing assays are not suited for the analysis of such isolates.