BACKGROUND: The Pall third-generation enhanced bacterial detection system (eBDS) was recently approved for detection of bacterial contamination in leukoreduced platelets (PLTs). The method is based on the measurement of the oxygen content as a marker for bacteria. eBDS incorporates major modifications including removal of the sample-set filter, modification of the culture medium, and incubation with agitation of the sample pouch. STUDY DESIGN AND METHODS: Ten whole blood-derived random-donor PLT units collected on Day 1 after donation and 10 single-donor apheresis PLT units were spiked with low levels of bacteria in three different blood transfusion centers. Inoculation was performed at a final concentration of 5 to 50 colony-forming units per mL with reference strains of five organisms involved in severe transfusion-associated infections. PLT units were stored at 22 degrees C for 24 hours before sampling. Six sample sets were then sterile-connected to each unit and placed on a horizontal agitator at 35 degrees C for 18 or 24 hours of incubation. RESULTS: No false-positive results were obtained, indicating a 100 percent specificity of the assay. Of 126 spiked sample pouches tested, 61 of 63 (96.82%) and 63 of 63 (100%) were detected positive after 18 or 24 hours of incubation, respectively. In the two missed cases that failed to detect Bacillus cereus, the measured oxygen was slightly above the detection threshold but was markedly different from the negative samples. CONCLUSION: The eBDS method allows definitive testing of PLTs as soon as 42 hours after collection and offers an alternative culture method to the BacT/ALERT system.