To assess the benefit of routinely used three-dimensional 1H-spectroscopy of the prostate combined with magnetic resonance imaging in patients with elevated prostate-specific antigen (PSA) levels and negative or no previous prostate biopsies. Fifty-four patients were examined with our combined imaging protocol, which consisted of transversal, coronal and sagittal T2-weighted fast spin echo sequences. For spectroscopy, we used a three-dimensional chemical shift imaging spin echo (3D-CSI-SE) sequence. The study population consisted of patients with elevated PSA levels and histologically proven prostate carcinoma and patients with elevated PSA levels and negative or no previous prostate biopsies. Examination time was 31 min, a time feasible for routine use. Eighty-eight tumour voxels and 67 control voxels of 27 patients with histologically proven prostate carcinoma were analysed. Ratios of (choline + creatine)/citrate [(Cho + Crea)/Cit] below 0.6 were classified as normal and above 0.6 as pathological. Applying this classification to 20 patients with tumour-suspicious lesions of the prostate and negative or no previous prostate biopsies, we could obtain a sensitivity and specificity for tumour detection of 100% and 69%, respectively. Our combined imaging protocol is feasible for routine use and can add valuable information for the diagnostic management of patients with negative or no previous prostate biopsies.