Phytoplasmas from the ribosomal subgroup 16SrXII-A (stolbur) were identified by PCR and RFLP analyses of 16S rRNA gene and elongation factor-Tu phytoplasma gene (tuf) in symptomatic grapevines from the Veles and Skopje areas in Macedonia. Two methods were used for nucleic acid extraction. Amplification and RFLP results of 16S rDNA were not influenced by the method used, but RFLP patterns of tuf gene revealed additional bands if a less time consuming method was applied. The possible origin of these bands is discussed. This is the first report of the presence of the stolbur phytoplasma in cvs Chardonnay and Vranac in Macedonia.