Background: Major depressive disorder (MDD) is a common mental disorder worldwide, but now there is a lack of clinically effective assessment and management of MDD. In this study, we used technetium-99 m ethylcysteinate dimer ([Tc-99m] ECD) SPECT/CT to characterize the regional cerebral blood flow (rCBF) status of MDD patients, and to explore an objective image assessment model of MDD which is non-or minimally-invasive, convenient and accurate in a clinical setting. Methods: The severity of MDD was assessed by three trained psychiatrists, based on scores obtained from HAMD and HAMA. [Tc-99m] ECD rCBF SPECT/CT was performed in 20 healthy controls and 74 unipolar MDD patients before receiving the treatment. The CT attenuation-corrected SPECT images data were automatically registered, analyzed simultaneously by 3D-SSP and eZIS. Results: The mean score of HAMD and HAMA in the MDD patients was 25.49 +/- 6.00, and 23.12 +/- 5.83, respectively. There was a positive correlation between two scores. The MDD women had higher HAMD scores than MDD men. The decreased rCBF of MDD patients in frontal lobes (bilateral B11, B47 and right B4, B6, B10, B46), temporal lobe (right B21, B41, B42) and cingulated cortex (bilateral B24, B33), while their increased rCBF in occipital lobe (bilateral B17, B19 and left B18). Additionally, the depression severity was negatively correlated with decreased rCBF in left ventral anterior cingulate cortex B24, and was positively correlated with decreased rCBF in left inferior prefrontal gyrus B47 and increased rCBF in right associative visual cortex B19. The anxiety severity was negatively correlated with decreased rCBF in left subgenual cortex B25. Conclusions: Although the mechanism underlying the correlation is not yet fully understood, our findings indicated that the rCBF SPECT/CT may provide an objective assessment for MDD severity. It might be used monitoring therapeutic efficacy in the management of MDD.