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When 155 ticks collected in different regions of Switzerland were tested by the hemolymph test, 10.3% were found to contain spotted fever group rickettsiae. Six rickettsial isolates were made from Dermacentor marginatus ticks and three were made from Ixodes ricinus ticks. The polymerase chain reaction followed by restriction fragment length polymorphism analysis showed that the Dermacentor ticks were infected with Rickettsia slovaca and the Ixodes ticks were infected with a spotted fever group rickettsia. Microimmunofluorescence serologic typing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins, and Western blot assay with polyclonal mouse antisera confirmed the results and determined that the Ixodes were infected with R. helvetica, the only previously described Swiss rickettsia. However, an additional new strain that could not be isolated was detected in one I. ricinus by hemolymph test and provisionally characterized by enzymatic restriction of its amplified DNA.