Recognizing persistent infection after resection arthroplasty and implantation of cement spacers in the infected total knee arthroplasty is often difficult. The purpose of this study was to determine whether aspiration of the knee after resection arthroplasty is valuable for determining the presence of ongoing infection. Thirty-four previously infected knees that were aspirated and cultured after resection arthroplasty, implantation of cement spacers, and intravenous antibiotics for an average of 6.3 weeks were identified. There were 8 cases of persistent infection-none identified on preoperative aspiration. Two preoperative cultures were false positive for Staphylococcus epidermidis. Preoperative cultures were negative in 32 knees, with 8 false-negative results. The antibiotic-free interval among all cases averaged 20 days; the cases with false-negative results from aspiration had an antibiotic-free interval averaging 11.5 days, compared with an average of 26 days among all other cases. Aspiration of knees after resection arthroplasty had sensitivity of 0%, positive predictive value of 0%, and accuracy of 71%. Specificity was 92%, and negative predictive value was 75%. A negative result from joint aspiration after resection arthroplasty does not necessarily rule out the presence of ongoing infection. False-negative results may be observed if joint aspiration is not delayed more than 2 to 3 weeks.