Background: The availability of a serologic test for cat scratch disease in humans has allowed the diagnosis of an increasing number of cases of this disease in disease in Chile. Aim: To perform a serological survey for Bartonella henselae among cats in Chile. Material and methods: Blood samples from 187 cats living in three Chilean cities were obtained. IgG and methods. Blood samples from 187 cats living in three Chilean, cities were obtained. IgG antibodies against Bartonella henselae were measured using indirect immunofluorescence. Blood cultures were clone in 60 samples. The presence of Bartonella henselae in positive cultures was confirmed by restriction fragment length polymorphism, polimerase chain reaction (RFLP-PCR). Results: The general prevalence of IgG antibodies against Bartonella henselae was 85.6%. No differences in this prevalence were formed among cats younger or older than 1 year, or those infested or not infested with fleas. However domestic cats had a lower prevalence when compared with stray cats (73 and 90% respectively, p <0.01). Bartonella henselae was isolated in 41% of blood cultures. All the isolated were confirmed as Bartonella henselae by RFLP-PCR. Conclusions: This study found all important reservoir of Bartonella henselae in Chilean cats and therefore a high risk of exposure in humans who have contact with them.