The epidemiology of infectious diseases depends on many characteristics of disease progression, as well as the consistency of these processes across hosts. Longitudinal studies of infection can thus inform disease monitoring and management, but can be challenging in wildlife, particularly for long-lived hosts and persistent infections. Numerous tortoise species of conservation concern can be infected by pathogenic mycoplasmas that cause a chronic upper respiratory tract disease (URTD). Yet, a lack of detailed data describing tortoise responses to mycoplasma infections obscures our understanding of URTDs role in host ecology. We therefore monitored Mycoplasma agassizii infections in 14 captive desert tortoises and characterised clinical signs of disease, infection intensity, pathogen shedding and antibody production for nearly 4 years after initial exposure to donor hosts. Persistent infections established in all exposed tortoises within 10 weeks, but hosts appeared to vary in resistance, which affected the patterns of pathogen shedding and apparent disease. Delays in host immune response and changes to clinical signs and infection intensity over time resulted in inconsistencies between diagnostic tools and changes in diagnostic accuracy throughout the study. We discuss the implications these results have for URTD epidemiology and past and future research assessing disease prevalence and dynamics in tortoise populations.