The choice of a diagnostic method depends on the characteristics of the herd to be analysed. Two herds with different prevalences of enzootic bovine leukaemia were chosen to study the concordance between agar gel immunodiffusion (AGID), enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) methods. PCR, an increasingly used virological method, was performed with four sets of primers, amplifying different genomic regions (env, pol and tax), from DNA extracted either from peripheral blood monocytes (PBMCs) or milk leucocytes. The highest percentage of positive animals was obtained using PCR performed with DNA extracted from PBMCs using primers which amplified either env or pol, followed by PCR using PBMCs and primers which hybridized with tax, then ELISA using serum and finally AGID. The results of PCR were more consistent with PBMCs than when milk leucocytes were used.