Detecting cancerous lesions is a major clinical application in emission tomography. In previous work, we have studied penalized maximum-likelihood (PML) image reconstruction for the detection task, where we used a multiview channelized Hotelling observer (mvCHO) to assess the lesion detectability in 3D images. It mimics the condition where a human observer examines three orthogonal views of a 3D image for lesion detection. We proposed a method to design a shift-variant quadratic penalty function to improve the detectability of lesions at unknown locations, and validated it using computer simulations. In this study we evaluated the bene t of the proposed penalty function for lesion detection using real data. A high-count real patient data with no identi able tumor inside the eld of view was used as the background data. A Na-22 point source was scanned in air at variable locations and the point source data were superimposed onto the patient data as arti cial lesions after being attenuated by the patient body. Independent Poisson noise was added to the high-count sinograms to generate 200 pairs of lesion-present and lesion-absent data sets, each mimicking a 5-minute scans. Lesion detectability was assessed using a multiview CHO and a human observer two alternative forced choice (2AFC) experiment. The results showed that the optimized penalty can improve lesion detection over the conventional quadratic penalty function.