Thirteen monoclonal antibodies (Mabs) specific for apple chlorotic leaf spot virus (ACLSV), produced by the somatic cell hybridization technique, were used to investigate the antigenic structure of the virus. Epitope specificity studies showed that these Mabs defined in ACLSV particles seven independent antigenic domains, representing at least eight distinct epitopes. One of them was present only in virions and not in dissociated subunits. It appeared that the interaction between a Mab and the virus could, in some cases, induce conformational changes in the viral particles which enhanced the binding of others. Twenty nine virus isolates differing in geographical origin, primary hosts and symptomatology were tested with these monoclonal antibodies by ELISA. With the exception of two Mabs which did not react with three cherry isolates, and one Mab which did not react with one plum isolate, all of them recognized all ACLSV isolates tested.