The aim of this study was to shed light on the association between lumbosacral transitional vertebra (LSTV) and degenerative lumbar spinal stenosis (DLSS). A cross-sectional retrospective study was performed on 165 individuals that were diagnosed with clinical picture of DLSS (age range: 40-88 years; sex ratio: 80M/85F) and 180 individuals without DLSS related symptoms (age range: 40-99 years; sex ratio: 90M/90F). All participants had undergone high-resolution CT scan for the lumbar region in the same position. We also used the volume rendering method to obtain three-dimensional CT images of the lumbosacral area. Both males and females in the stenosis group manifest greater prevalence of LSTV than their counterparts in the control group (P<0.001). Furthermore, the presence of LSTV increases the likelihood of degenerative spinal stenosis (odds ratio= 3.741, P<0.001). In the control group, LSTV was more common in males, and sacral slope angle of males was significantly greater in LSTV group compared to non-LSTV. This study indicates that LSTV was significantly associated with symptomatic DLSS.