ABSTRACTBacteria of the genusBartonella, a member of theAlphaproteobacteria, are fastidious, Gram-negative, aerobic bacilli that comprise numerous species, subspecies, and genotypes.Bartonella henselae, with a worldwide distribution, infects cats, dogs, horses, humans, and other mammals. Diagnostically, direct detection ofBartonella henselaein patient blood specimens by culture or molecular methods is required to confirm infection with this bacterium. Enrichment blood culture combined with quantitative PCR (qPCR) or ddPCR enhances the sensitivity of direct detection. The addition of sheep blood to liquid culture media increased theBartonella henselaeDNA concentration compared to controls, additionally improving PCR direct detection sensitivity.IMPORTANCEThis study aims to improve diagnostic detection ofBartonella henselae. Patient samples are combined with enriched bacterial cultures aimed at growingBartonella henselaefor the best possible chance at detection. However, currentBartonellagrowth methods could be improved. The DNA extraction method used by most laboratories should also be optimized. Sheep blood was added to increase the growth ofBartonella henselaeand multiple DNA extraction methods were to be compared to each other.