The genetic diversity of Grapevine fanleaf virus (GFLV) was evaluated in 4 isolates sampled from naturally infected grapevines from South Moravia (Czech Republic) and 2 Italian isolates from Bari (Italy). Conserved regions within sequences in databases were found and new primers corresponding to these regions were designed and tested for RT-PCR amplification of the CP codifying region. After sequencing of obtained amplicons the similarity of isolates was analysed via alignments of sequences and by means of dendrograms.