Pulmonary disease is by far the most frequent disease caused by nontuberculous mycobacteria (NTM). To diagnose NTM pulmonary disease (NTM-PD), patients should have symptoms and radiologic signs suggestive of NTM-PD, and cultures of multiple respiratory tract samples must grow the same NTM species. Thus, the microbiological laboratory has a central role in the diagnosis of NTM-PD. This review summarizes currently available data on techniques involved in the microbiological diagnosis of NTM-PD, and aims to provide a framework for optimal microbiological diagnosis. (Source: Clinics in Chest Medicine)