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To evaluate the usefulness of a multiplex polymerase chain reaction (PCR) assay as a complementary tool in the diagnosis of prosthetic joint infections in the routine setting of a clinical microbiology laboratory, with a special focus on patients at high risk of culture-negative infections and high suspicion of infection. The results obtained in the routine care setting with the use of the commercial multiplex PCR (Unyvero i60©, Curetis AG, Holzgerlingen, Germany) were retrospectively reviewed. The test was performed in samples of pati