In the present study, the determination of the presence of Bartonella henselae in Istanbul in 96 pet, shelter and stray cats were aimed. The samples from the cats were examined by two different bacteriologic culture methods. The blood samples collected into pediatric lysis isolator tubes were spun and the pellet inoculated onto Heart Infusion Agar supplemented with 5% rabbit blood. BACTEC Peds Plus/F were inoculated with samples and incubated in the BACTEC 9050 automated blood culture systems. After the incubation time, growth were evaluated according to the colony morphology, growth time, gram properties and biochemical properties, and it was determined that 27 isolates by solid medium, 10 isolates by BACTEC were Bartonella spp. Isolates were determined as Bartonella henselae by PCR amplification using specific primers of a portion of 16S-23S rRNA intergenic region. In conclusion, the prevalence of Bartonella henselae bacteremia was found 28.1% of cats in Istanbul.