To detect non-bacteriuric renal infection in a diabetic patient, though difficult but is very important because early diagnosis and early treatment can prevent later complications such as renal abscess, renal hypertension or even end stage renal disease. Herein, we presented a case of diabetic patient with septicemia whose urine culture and renal ultrasonography were negative initially. By using a combination of dual isotope images and single photon emission computed tomography technique, an infectious lesion in the upper pole of left kidney was revealed, which was identified as acute focal bacterial nephritis by computed tomography four days later. This case report showed that 67Ga plus 99mTc dimercaptosuccinic acid images are useful in patients under clinical suspicion of renal infection, especially for those with negative urine analysis and/or urine culture initially.