Objective To summarize the best evidence of sputum collection methods for patients with pulmonary tuberculosis. Methods We searched websites and databases including, but not limited to, BMJ best practice, UpToDate, World Health Organization, Registered Nurses’ Association of Ontario, National Institute for Health and Care Excellence, Guidelines International Network, Infectious Diseases Society of America,Cochrane Library,CINAHL(EBSCO), EMBASE(OVID), PubMed, Web of Science, JBI, CNKI, Wanfang, VIP, and CBM. Guidelines,recommended practices,evidence summaries,expert consensuses, and systematic reviews regarding collection methods of sputum samples for patients with pulmonary were considered. 2 researchers independently assessed methodological quality of included papers and extracted data. Results A total of 13 papers were identified including 6 guidelines, 3 systematic reviews,and 4 expert consensuses. 24 pieces of evidence were categorized into 6 aspects including pre-collection evaluation,pre-collection intervention,collection methods,sputum sample evaluation and transportation,occupational protection, and patient education. Conclusion This evidence summary provided sufficient evidence-based information for healthcare professionals to conduct sputum specimen collection for patients with pulmonary tuberculosis. Healthcare professionals should also consider feasibility, appropriateness, meaningfulness, and effectiveness of evidence in local context before adopting a certain method of sputum collection.