Analysis of 483 skeletons from Arica (Chile) and review of mummy dissection records demonstrates an overall 1% prevalence rate for tuberculosis between 2000 B.C. and A.D. 1500. Tuberculosis cases cluster in the period A.D. 500–1000 which correlates with fully agropastoral societies. Considering only these agropastoral societies, about 2% of their members show tuberculosis lesions. A segment of DNA unique to Mycobacterium tuberculosis was identified in an extract from the vertebral lesion of a 12-year-old girl with Pott's disease from about A.D. 1000, establishing the pre-Columbian presence of tuberculosis with the most specific evidence currently available. © Wiley-Liss, Inc.