Background: Pain affected gait performance in patients with non-specific chronic low back pain (NSCLBP) due to protective mechanisms in response to pain. Insufficient evidence demonstrated whether pain altered the walking patterns in NSCLBP. Therefore, this review aimed to compare the walking patterns between individuals with and without NSCLBP. Methods: The literature was systematically reviewed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The Cochrane, PubMed, Web of Science (WOS), EMBASE, and EBSCOhost were searched for the relevant studies, which were included if they: 1) compared patients with NSCLBP to the healthy controls, 2) evaluated spatiotemporal gait characteristics, 3) were published in English, 4) were peer-reviewed original research articles, and 5) were published since the year of 2000. Two researchers independently assessed the quality of the studies using the modified Downs and Black checklist. Meta-analyses with the random effects model were used, and the standardized mean differences (SMD) and the 95% confidence intervals (CI) were analyzed. Results: Out of 822 records, thirteen studies were included in the systematic review, and twelve were incorporated into the meta-analysis. The meta-analysis revealed significant differences between NSCLBP patients and healthy controls in walking speed (SMD = –0.67, 95% CI: –1.08 to –0.25, p = 0.002, I2 = 76%) and step length (SMD = –0.55, 95% CI: –0.94 to –0.17, p = 0.004, I2 = 64%). Discussion: Within spatiotemporal gait parameters, patients with NSCLBP show moderately consistent reductions in walking speed and step length relative to healthy controls, which may reflect pain-related protective strategies. These alterations underscore clinically meaningful gait impairments that should be considered in the design of targeted rehabilitation programs. PROSPERO registration number is CRD42023418511.