Of the 39 aspirations in 32 patients included in the study, there were 33 (85%) knee and 6 (15%) hip samples. Eleven (28%) samples met MSIS criteria for PJI. Of the 23 samples that had recent antibiotic use (6 MSIS positive, 17 negative), alpha-defensin results confirmed the correct MSIS diagnosis in 19 (83%) samples. Of the 11 samples in which alpha-defensin testing was performed for a borderline cell count (3 MSIS positive, 8 negative), alpha-defensin confirmed the MSIS diagnosis in 10 of 11 (91%) samples. Finally, among the 5 samples with suspected false-positive or false-negative cultures (2 MSIS positive, 3 negative), alpha-defensin confirmed the correct diagnosis in 3 (60%) samples. The overall sensitivity, specificity, negative predictive value, and positive predictive value of synovial alpha-defensin were 82%, 82%, 92%, and 64%, respectively.