This is a case study of a 23 year old male diagnosed with Acanthamoeba keratitis. Initial misdiagnosis and inappropriate treatment lead to increased severity of the infection requiring surgical intervention. This case illustrates the complexity of Acanthamoeba keratitis infection, highlighting the diagnostic challenges which often begin with misdiagnosis and treatment, the role of the laboratory in providing confirmatory testing, and the relevance of patient knowledge related to contact lens wear and care.