Abstract Non-tuberculous mycobacteria (NTM) comprise a large, diverse group of bacteria usually found in soil, water and wild and domestic animal populations. NTM were not recognized as significant sources of pulmonary disease until relatively recently. Unlike Mycobacterium tuberculosis, NTM do not cause tuberculosis (TB), but can cause skin and soft tissue infections, disseminated infections and severe lung disease, and pose a particular threat for those afflicted with cystic fibrosis, chronic obstructive pulmonary disease (COPD) or pre-existing bronchiectasis. There is currently no vaccine to protect from NTM infections. NTM are also intrinsically resistant to most existing antimicrobials. Among new antimicrobials currently under various stages of development, including those to treat TB, few have potent anti-NTM activity. Multiple studies indicate that the prevalence of NTM disease is increasing globally. The incidence and prevalence of NTM lung disease (NTM-LD) in the U.S. is also increasing, and now surpasses that of TB. However, we have a fundamentally poor understanding of NTM and the diseases they cause. The goals of the Colorado Mycobacteria Conference: Focus on NTM are to help fill the existing gaps in basic and translational NTM research by bringing together junior and senior scientists, clinicians and public health officials that do not often attend the same conferences, highlighting new findings, discussing lessons learned and, importantly, training the next generations of researchers and clinicians.