[Objective] Explore the role of Dithiothreitol in bacterial detection in Prosthetic Joint Infection (PJI) patients. [Methods] In a prospective study, three different treatments (A: no additional treatment; B: periprosthetic tissues were treated with isotonic saline for 15 minutes; C: periprosthetic tissues were treated with 1mg/ml DTT for 15 minutes) were performed to periprosthetic tissues from 46 patients presented with chronic PJI in our hospital and cultured for bacterial detection. Bacterial detection rate were compared among the three different treatments in these 46 patients and those who undertook antibiotics treatment 2-4 weeks prior to samples collected in our department. [Results]When compared among the 46 patients, bacterial detection rate in treatment C is 69.57%（32/46）much higher than in A 43.48%（20/46）and B 36.96%（17/46）, and the difference is significant. However, when compared among 28 patients who undertook antibiotics treatment 2-4 weeks prior to samples collected, although bacterial detection rate in treatment C is 57.00%（16/28）higher than in A 36.00%（10/28）and B 28.57%（8/28）, the difference is not significant. [Conclusion] DTT can improve the bacterial detection rate in chronic PJI patients.