Acanthamoeba keratitis is a rare, serious protozoal infection of the cornea associated with wearing contact lenses. To identify risk factors in soft contact lens wearers, we interviewed 27 patients with Acanthamoeba keratitis and 81 uninfected matched controls to compare contact lens care practices, brands of contact lenses and associated solutions, and behavioral activities. Patients were significantly more likely than controls to use homemade saline instead of commercially prepared saline (21/27 [78%] vs 14/81 [17%]; odds ratio [OR], infinity), and wear their lenses while swimming (17/27 [63%] vs 24/81 [30%]; OR, 6.2). Contact lens disinfection schedules could be determined for 25 of the patients and all of the controls. Patients were significantly more likely than controls to disinfect their lenses less frequently than recommended by lens manufacturers (18/25 [72%] vs 26/81 [32%]; OR, 5.8). Microbiologic assay of contact lens solutions from controls showed frequent contamination with high levels of bacteria. Acanthamoeba species were isolated from homemade saline solutions from two controls. These findings emphasize adherence to recommended methods of soft contact lens care, especially when using nonsterile lens care solutions.