Previous studies showed that cerebral small vessel disease (cSVD) is a leading cause of cognitive decline in elderly people and the development of Alzheimer's disease. Although brain structural changes of cSVD have been documented well, it remains unclear about the properties of brain intrinsic spontaneous activity in patients with cSVD.We collected resting-state fMRI (rs-fMRI) and T1-weighted 3D high-resolution brain structural images from 41 cSVD patients and 32 healthy controls (HC). By estimating the amplitude of low-frequency fluctuation (ALFF) under three different frequency bands (typical band: 0.01-0.1 Hz; slow-4: 0.027-0.073 Hz; and slow-5: 0.01-0.027 Hz) in the whole-brain, we analyzed band-specific ALFF differences between the cSVD patients and controls.The cSVD patients showed uniformly lower ALFF than the healthy controls in the typical and slow-4 bands (pFWE < 0.05). In the typical band, cSVD patients showed lower ALFF involving voxels of the fusiform, hippocampus, inferior occipital cortex, middle occipital cortex, insula, inferior frontal cortex, rolandic operculum, and cerebellum compared with the controls. In the slow-4 band, cSVD patients showed lower ALFF involving voxels of the cerebellum, hippocampus, occipital, and fusiform compared with the controls. However, there is no significant between-group difference of ALFF in the slow-5 band. Moreover, we found significant "group × frequency" interactions in the left precuneus.Our results suggested that brain intrinsic spontaneous activity of cSVD patients was abnormal and showed a frequency-specific characteristic. The ALFF in the slow-4 band may be more sensitive to detecting a malfunction in cSVD patients.Copyright © 2023 Chen, Huang, Zhang, Huang, Ling, Liu and Yang.