Background Bacterial infection can pose a substantial diagnostic dilemma. Techniques involving radiolabelled leucocytes can pinpoint the site of inflammation. However, previous radiolabelling techniques have failed to distinguish between bacterial-mediated infection and non-bacterial inflammation. To overcome this difficulty, we have studied a radiopharmaceutical, technetium-99m (99mTc) Infecton, which is based on the antibiotic ciprofloxacin. Methods We used this agent to image bacterial infection in 56 patients (one twice) with known or suspected sites of infection. We then compared the imaging results of these patients with those from a radiolabelled leucocyte study. Findings The concordance rate was 68% (39 out of 57 images). In 18 discordant results 99mTc Infecton was correctly positive in 8 out of 9 positive studies and correctly negative in 4 out of 9 negative studies. 4 out of 5 of the falsely negative studies were in patients who had taken antibiotics for over 7 days. We found that 99mTc Infecton gave better imaging results than radiolabelled leucocytes. Comparison between 99mTc Infecton and leucocyte imaging gave sensitivities of 84% and 81%, and specificities of 96% and 77%, respectively. Interpretation We believe that the specificity 99mTc Infecton confers for bacterial infection and its ease of administration are the main advantages of this new agent.