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To assess the performance of the rapid syndromic BioFire® Joint Infection Panel (BF-JIP) to detect bacterial and fungal pathogens, as well as antibiotic resistance genes, directly in synovial fluid specimens collected from patients with acute arthritis. The study was conducted in six French bacteriological laboratories. To assess the performances of BF-JIP, results were compared with those of synovial fluid 14-day culture and, in case of discrepancy, with those of complementary molecular methods and intraoperative samples. A total of 308 synovial fluid specimens were tested after collection from 308 adults and children presenting with clinical and biological suspicion of acute arthritis; patients presenting with acute periprosthetic joint infection were included according to the European Bone and Joint Infection Society 2021 criteria. Only one specimen failed (no result). On the basis of the consolidated data, the BF-JIP was concordant with the 14-day culture in 280 (91.2%) of the 307 specimens finally included in the study. The positive percentage agreement was 84.9% (95% CI, 78.8–89.8%) and the negative percentage agreement was 100% (95% CI, 97.2–100%). The positive predictive value was extremely high (100%; 95% CI, 97.6–100%), whereas the negative predictive value was lower (82.6%; 95% CI, 75.7–88.2%), partially explained by the missing target species in the panel. The BF-JIP showed high performances to detect pathogens involved in acute arthritis. [ABSTRACT FROM AUTHOR]