This method was aimed targeting more Campylobacter species than conventional PCR-based identifications. They generally use species-specific primers focusing on clinically common species like C. jejuni, resulting in failure to recognize other species. We made the PCR-based identification more flexible using degenerate primers and DdeI- and MboI-separately used RFLP assay, which were designed on the basis of gyrB nucleotide sequence data of 14 Campylobacter species including C. jejuni, C. coli, and C. fetus. Ninety-four clinical isolates from patients with Campylobacter gastroenteritis and 13 biochemically identified C. fetus were used for its evaluation. In consequence, this method succeeded in identifying C. jejuni, C. coli, and C. fetus with tentative sensitivity (93.4-98.0%) and specificity (89.0-99.0%). According to our data-based analysis, the primers can possibly target other related species including Helicobacter and Arcobacter. This method may be a universal identification for Campylobacter and related organisms and would provide an alternative identification in clinical microbiology.