This study reported four cases of disseminated non-tuberculous mycobacterial (NTM) infection caused by local puncture, and elaborated the pathway of infection, their clinical manifestations, the process of diagnosis and treatment. The study aimed to improve the clinicians’ recognition on the sterilization of acupuncture, and prevent the iatrogenic infection. Both the 4 cases had a history of local puncture, and diagnosed as disseminated NTM infection. They recovered to a better situation after receiving the anti- NTM infection treatment. Recently, there were few reports about disseminated NTM infection caused by local puncture. The clinical manifestation is not obvious, it usually misdiagnosed as tuberculosis mycobacteria. Thus, it’s necessary to improve the recognition of infection routes and clinical manifestations of NTM, and establish a standard operational disinfection procedure.