DNA of Treponema pallidum subsp. pallidum Nichols strain was compared to DNA isolated from three different strains of T. pallidum subsp. pertenue (strain Gauthier, Samoan D, CDC-2). As a result of DNA microarray comparisons, 25 genes (13 with stronger and 6 with weaker signal in pertenue strains and 6 control genes with similar signal in both subspecies examined) were selected and sequenced. Altogether, 24083 nucleotides (2.12% of the genome) were sequenced in 3 pertenue strains and control Nichols. No region of extensive sequence heterogeneity was detected. However, 15 different single nucleotide polymorfisms (SNP) were identified: 3 SNPs in Gauthier strain, 14 in Samoan D and 15 SNPs in CDC-2. Ten (out of 15) SNPs cause amino acid changes. SNPs common for all pertenue strains as well as SNPs specific for each individual strain will allow to use these nucleotide polymorfisms to design sequence specific PCR diagnostics of these strains.