The interaction of the terbium-difloxacin complex (Tb-DFX) with DNA has been examined by using UV-vis absorption and luminescence spectroscopy. The Tb-DFX complex shows an up to 85-fold enhancement of luminescence intensity upon titration with DNA. The long decay times allow additional detection schemes like time-resolved measurements in microplate readers to enhance sensitivity by off-gating short-lived background luminescence. Optimal conditions are found at equimolar concentrations of Tb(3+) and DFX (0.1 or 1 microM) at pH 7.4. Under these conditions, the luminescence intensity is linearly dependent on the concentration of ds-DNAs and ss-DNA between 1-1500 ng mL(-1) and 4.5-270 ng mL(-1), respectively. The detection limit is 0.5 ng mL(-1) for ds-DNAs and 2 ng mL(-1) for ss-DNA. The mechanism for the luminescence enhancement was also studied.