The specificity of serological tests for Lyme borreliosis is impaired by cross-reacting antibodies. In order to select antigens for more specific tests, specific and cross-reactive proteins of Borrelia burgdorferi must be identified. Therefore, to analyze cross reactions of Borrelia burgdorferi with other bacteria, rabbit immune sera against heterologous bacteria (Borrelia hermsii, Treponema pallidum, Treponema phagedenis, Leptospira interrogans (serogroup grippotyphosa), Neisseria meningitidis, Haemophilus influenzae, Yersinia enterocolitica (serotypes O3 and O9), Campylobacter jejuni, Listeria monocytogenes O1, Pseudomonas aeruginosa, Escherichia coli, Salmonella typhimurium, Shigella flexneri and Legionella micdadei) were examined by Western blot using Borrelia burgdorferi as antigen. Broad cross reactivity was shown for Borrelia proteins of the 60-75 kDa range. Other broadly cross-reacting proteins were at the level of p40, p33 and two proteins in the range of 20 kDa. Some of the cross reactions were eliminated by absorption of the sera with Treponema phagedenis. The absorbed antibodies were directed mainly against bands at the level of p33 and bands of the 60 to 75 kDa range. Showing the lowest potential for cross reactivity, p100, p41, OspA and pC seem to be the most suitable antigens for serodiagnosis. In contrast to p100 and OspA, however, p41 and pC showed cross reactivity with immune sera against bacteria not belonging to the genus Borrelia.