Objective To provide information regarding the procedures, safety, tolerability, and measurement properties of the 6-min step test. Data sources MEDLINE, EMBASE, CINAHL, and SPORTDiscus (from inception until January 2024). Review methods Studies that examined adults with acute or chronic diseases, and outcomes related to procedures, safety, tolerability, or measurement properties of the 6-min step test were included. Outcome data were summarized and combined in meta-analyses. The quality of included studies was assessed by the Consensus-based Standards for the selection of health Measurement Instruments checklist, and the quality of evidence was determined according to the Grading of Recommendations Assessment, Development, and Evaluation system. Results Fourteen studies, involving 847 participants, were included. All studies performed the 6-min step test in 6 min; however, some studies varied the step height and the use of upper limb support. The test appears to be safe and well tolerated by individuals. Moderate- to high-quality evidence demonstrated appropriate results for test-retest reliability (4 studies; Intraclass correlation coefficient 0.96; 95% CI 0.91–0.98; n = 125), criterion validity (4 studies; r = 0.53; 95% CI 0.30–0.71; n = 307), and construct validity (4 studies; r = 0.63; 95% CI 0.52–0.73; n = 233). Conclusion This review provides recommendations for applying the 6-min step test in clinical and research settings. No adverse events were reported, and the test appears to be well tolerated. Adequate results were found for test-retest reliability, criterion validity, and construct validity. Review Registration PROSPERO (CRD42022347744).