Enzymatic DNA amplification was applied to DNA and elementary bodies of C. trachomatis. Oligonucleotide primers were chosen in a sequence of a conserved domain of the major outer membrane protein to generate the amplification of a 129-base pair fragment. This sequence was amplified in the 15 serovars of C. trachomatis; however, serovar J gave a weaker signal than the others. The specificity was controlled by EcoRI restriction enzyme digestion and Southern analysis using an internal probe of the amplified sequence. No cross-reaction was shown with DNA of 11 other bacteria. Thus, enzymatic DNA amplification by the polymerase chain reaction appears to be a potential tool for the specific detection of C. trachomatis.