Objective To retrospectively study the distribution of bacterial species, clinical characteristics, as well as diagnosis and treatment information of non-tuberculous Mycobacterium (NTM) infections in bone and joints in a tertiary orthopedic hospital over the past five years, providing reference for clinical diagnosis, treatment, and rela-ted research of NTM infections in bone and joints. Methods Clinical data of patients diagnosed with NTM infections in bone and joint systems from January 2016 to December 2020 were collected. Cases that met the criteria were selected. Distribution of bacterial species, basic characteristics, clinical manifestations, treatment and prognosis information of patients with different types of infections were summarized and analyzed. Results From 2016 to 2020, a total of 24 cases of NTM infections in bone and joints were diagnosed. Among them, Mycobacterium fortuitum was found in 10 cases, Mycobacterium abscessus in 6 cases, Mycobacterium chelonae-Mycobacterium abscessus group in 3 cases, Mycobacterium houstonense in 3 cases, Mycobacterium smegmatis in 1 case, and Mycobacterium avium in 1 case. 13 patients had underlying diseases of varying degrees, and the infected sites involved various joints in the limbs, with the knee joint (n=9) and ankle joint (n=7) being the most common. All 24 patients presented abscesses and tenderness at the infected site, with a few cases showing local skin temperature elevation or systemic fever. 50.0% (12/24) of patients had sinus formation, and 20.8% (5/24) had granulomas. Among the 24 patients, 2 did not undergo surgery, while the other 22 underwent surgical treatment. All patients received both internal and external adjuvant treatment with antimicrobial agents combined with traditional Chinese medicine, with a median cure duration of 129 days and a cure rate of 83.3% (n=20). One patient was transferred to other hospital after pathogen detection, one died due to multiple organ failure, and two had poor treatment outcomes. Conclusion Timely and accurate early pathogenic diagnosis can shorten the diagnostic period of NTM infections in bone and joints, enabling timely, targeted and precise treatment to effectively reduce infection-related damage, and alleviate patient pain. Traditional Chinese medicine-assisted treatment can promote wound healing, shorten treatment course, and reduce adverse reactions.