An outbreak of Acanthamoeba keratitis, a rare, potentially blinding, corneal infection, was detected in the United States in 2007; cases had been increasing since 2004. A case-control study was conducted to investigate the outbreak. We interviewed 105 case-patients from 30 states and 184 controls matched geographically and by contact lens use. Available contact lenses, cases, solutions, and corneal specimens from case-patients were cultured and tested by molecular methods. In multivariate analyses, case-patients had significantly greater odds of having used Advanced Medical Optics Complete Moisture Plus (AMOCMP) solution (odds ratio 16.9, 95% confidence interval 4.8-59.5). AMOCMP manufacturing lot information was available for 22 case-patients, but none of the lots were identical. Three unopened bottles of AMOCMP tested negative for Acanthamoeba spp. Our findings suggest that the solution was not intrinsically contaminated and that its anti-Acanthamoeba efficacy was likely insufficient. Premarket standardized testing of contact lens solutions for activity against Acanthamoeba spp. is warranted.