We conducted a serosurvey on Mediterranean spotted fever (MSF), in a nonendemic area using western blot and microimmunofluorescence. Among 262 tested sera, 53 were positive by micro-immunofluorescence at a titer of 50. When 48 positive sera were western blot tested, 15 did not exhibited any reaction, 17 reacted against the non-specific lipopolysaccharide, and only 16 reacted against the specific protein antigens. Fourteen of the sera with a specific reaction were sampled in a village with a unique submediterranean climate. Western blot may be a more specific tool to determine the real seroprevalence of MSF.