Present guidelines recommend culturing only central venous catheter (CVC) tips from patients with suspected catheter-related bloodstream infection (CR-BSI). However, a high proportion of these suspicions are not confirmed. Moreover, CVC tip culture increases laboratory workload, and reports of colonization may be meaningless or misleading for the clinician. Our working hypothesis was that CVC tips should be refrigerated and cultured only in patients with positive blood cultures. We evaluated the effect of 6-day refrigeration of 215 CVC tips. We selected all the catheters with a significant count according to the Maki's roll-plate technique and randomly assigned them to 2 groups. In group A, the catheters were recultured after 24 h of refrigeration, and in group B, the catheters were recultured after 6 days more of refrigeration, so that the refrigeration time evaluated would be of 6 days. The yield of refrigerated CVC tips that grow significant colony counts of primary culture in group B was compared with the yield of refrigerated catheter tips in group A. The difference showed that 6-day refrigeration reduced the number of significant CVCs by 15.2%. Only 61 CVCs were obtained from patients with CR-BSI, and in most of them, blood cultures were already positive before CVC culture, so only 0.91% of the CR-BSI episodes would have been misdiagnosed as culture negative after refrigeration. Refrigeration of CVC tips sent for culture and culturing only those from patients with positive blood cultures reduce the workload in the microbiology laboratory without misdiagnosing CR-BSI.