(1) Background: Marker-based 3D motion capture systems (MBS) are considered the gold standard in gait analysis. However, they have limitations for which markerless camera-based 3D motion capture systems (MCBS) could provide a solution. The aim of this systematic review and meta-analysis is to compare the accuracy, validity, and reliability of MCBS and MBS. (2) Methods: A total of 2047 papers were systematically searched according to PRISMA guidelines on 7 February 2024, in two different databases: Pubmed (1339) and WoS (708). The COSMIN-tool and EBRO guidelines were used to assess risk of bias and level of evidence. (3) Results: After full text screening, 22 papers were included. Spatiotemporal parameters showed overall good to excellent accuracy, validity, and reliability. For kinematic variables, hip and knee showed moderate to excellent agreement between the systems, while for the ankle joint, poor concurrent validity and reliability were measured. The accuracy and concurrent validity of walking speed were considered excellent in all cases, with only a small bias. The meta-analysis of the inter-rater reliability and concurrent validity of walking speed, step time, and step length resulted in a good-to-excellent intraclass correlation coefficient (ICC) (0.81; 0.98). (4) Discussion and conclusions: MCBS are comparable in terms of accuracy, concurrent validity, and reliability to MBS in spatiotemporal parameters. Additionally, kinematic parameters for hip and knee in the sagittal plane are considered most valid and reliable but lack valid and accurate measurement outcomes in transverse and frontal planes. Customization and standardization of methodological procedures are necessary for future research to adequately compare protocols in clinical settings, with more attention to patient populations.
