Diagnosis of Propionibacterium acnes bone and joint infection is challenging due to the long cultivation time of up to 14 days. We retrospectively studied whether reducing the cultivation time to 7 days allows accurate diagnosis without losing sensitivity. We identified patients with at least one positive P. acnes sample between 2005 and 2015 and grouped them into "infection" and "no infection." An infection was defined when at least two samples from the same case were positive. Clinical and microbiological data, including time to positivity for different cultivation methods, were recorded. We found 70 cases of proven P. acnes infection with a significant faster median time to positivity of 6 days (range, 2 to 11 days) compared to 9 days in 47 cases with P. acnes identified as a contamination (P < 0.0001). In 15 of 70 (21.4%) patients with an infection, tissue samples were positive after day 7 and in 6 patients (8.6%) after day 10 when a blind subculture of the thioglycolate broth was performed. The highest sensitivity was detected for thioglycolate broth (66.3%) and the best positive predictive values for anaerobic agar plates (96.5%). A prolonged transportation time from the operating theater to the microbiological laboratory did not influence time to positivity of P. acnes growth. By reducing the cultivation time to 7 days, false-negative diagnoses would increase by 21.4%; thus, we recommend that biopsy specimens from bone and joint infections be cultivated to detect P. acnes for 10 days with a blind subculture at the end. P ropionibacterium acnes is a facultative anaerobic Gram-positive rod, abundant on the human skin, and mainly associated with the sebaceous glands of the shoulder and axilla (1). It is most commonly associated with the chronic skin disease acne vulgaris. However, it may also cause bone and joint infections, including implant-associated infections. P. acnes has been recognized as an emerging cause of shoulder infections (2, 3) and is among the most common pathogens isolated in shoulder periprosthetic joint infections (PJI) (4, 5). P. acnes has also been implicated in other biofilm-related infections (6-8), such as cardiovascular implantassociated infections (9), spinal osteomyelitis (10, 11), and endophthalmitis (12, 13).Diagnosis of P. acnes bone and joint infections is challenging since pain is often the only symptom (14, 15). For a long time, P. acnes was underdiagnosed in bone and joint infections due to the short cultivation time routinely used in diagnostic laboratories. In general, biofilm-forming bacteria are known to replicate at a slow rate due to low metabolism (16). Since recent studies recommended a prolonged cultivation time of up to 14 days for bone and joint infections (17, 18), the diagnosis of P. acnes infections has become more frequently documented (19). In view of the high costs of a long incubation period, a recent study suggested that 7 days of incubation should be sufficient for accurately diagnosing orthopedic implant-associated infections (20). In this study, 96.6% ...