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The aim of this study was to investigate the presence of spontaneous retinal venous pulsation (SVP) in patients with unilateral primary open-angle glaucoma (POAG) and low intraocular pressure (IOP).The SVP of 93 POAG patients with unilateral glaucoma and untreated IOP of ≤21 mm Hg was assessed using the movie tool of a confocal scanning laser ophthalmoscope (Spectralis HRA; Heidelberg Engineering). The frequency of SVP was compared between the glaucomatous and fellow eyes. Intereye differences in the frequency of SVP were assessed using McNemar test. A linear mixed-effect model was used to determine the factors associated with glaucomatous eyes, taking into account clustering of eyes within subjects.Forty-five patients had SVP in both eyes and 15 had SVP in neither eye. Of the remaining 33 patients who showed SVP only in 1 eye, 31 had SVP only in the fellow eye, and 2 had SVP only in the glaucomatous eye. The SVP was significantly less prevalent in glaucomatous eyes (50.5%) than in fellow control eyes (81.7%) (P<0.001). In the linear mixed-effect model, the presence of SVP (P<0.001) and higher untreated IOP (P=0.001) were the significant predictors for glaucoma.SVP was less frequently found in glaucomatous eyes than healthy fellow eyes in unilateral POAG patients with low IOP.