A rapid screening test (45 min) for bacteriuria was evaluated in 1,000 clinical urine specimens. The test procedure is based upon firefly luciferase analysis of bacterial ATP and uses the Lumac kit and Lumac M2010 Biocounter (3M Co., St. Paul, Minn.). The procedure allows for removal and destruction of nonbacterial ATP and subsequent analysis of bacterial ATP by firefly luciferase with a single photon counter. Results, expressed in relative light units, were compared with actual CFU by the calibrated loop technique. Sensitivities and specificities were calculated separately for clean-catch midstream specimens and for urines obtained by catheterization. The sensitivity for 719 clean-catch specimens with a Lumac cutoff of 2500 relative light units, representing 2 105 CFU/ml, was 93%. The sensitivity for 281 catheterized specimens with a Lumac cutoff of 2200 relative light units, representing i104 CFU/ml, was 95%. There were 19 false-negative results in the 1,000 specimens tested; more than 50% of these were contaminated cultures and were not considered significant in determining bacteriuria. In conclusion, the Lumac bioluminescence assay is a reliable, rapid bacteriuria screening technique with the potential of reducing the laboratory cost and for reducing the turnaround time in processing negative urine cultures.