7-Bromo-1,4-dihydro-4-oxo-quinolin-3-carboxylic acid (BDOQCA), was synthesized with a yield of 93% and well characterized. The obtained compound was investigated to label with one of the most important radioactive isotopes (technetium-99m). Effect of BDOQCA concentration, stannous chloride dihydrates (SnCl2.2H2O) concentration, pH and reaction time on the percent labeling yield of 99mTc-BDOQCA complex was studied in details. 99mTc-BDOQCA complex was obtained at a maximum yield of 97.3% by mixing 2.5 mg of BDOQCA with 25 μg SnCl2.2H2O at pH 6 and 30 min reaction time and the formed complex was stable for a time up to 8 h with a maximum yield of 97.3%. Biodistribution studies in mice were carried out using experimentally induced infection in the left thigh using E. coli. Both thighs of the mice were dissected and counted to evaluate the ratio of bacterial infected thigh/contralateral thigh. Higher uptake in the infected thigh was observed after 2 h of IV administration of 99mTc-BDOQCA complex (T/NT = 7.6 ± 0.6%) than that of the commercially available 99mTc-ciprofloxacin complex (T/NT = 3.8 ± 1%). The in vitro binding and biodistribution of 99mTc-BDOQCA complex in the septic and aseptic inflammation bearing mice showed that, 99mTc-BDOQCA complex is a promising agent for infection imaging and can differentiate between infected and inflamed muscle.