The aim of this study was to compare the antimicrobial action of commercially available multipurpose disinfection solutions (MPDS) with and without hydrogel contact lens for disinfection of isolated corneal bacteria. Methods: Five commercially available MPDS in Brazil (BioSoak, Clear Lens, OPTI-FREE, Renu, and UltraSept) were tested against Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis. All five MPDS were also tested on P. aeruginosa and S. aureus biofilm in a Hioxifilcon A lens. Results: OPTI-FREE and Renu were effective against all bacterial isolates without contact lenses. BioSoak was effective in inhibiting P. aeruginosa and S. epidermidis but not against S. aureus. UltraSept was effective for inhibiting S. epidermidis and S. aureus but not against P. aeruginosa. Clear Lens was effective in inhibiting only S. epidermidis but not P. aeruginosa and S. aureus. In contact lens bacterial biofilms, OPTI-FREE was the only MPDS to demonstrate significantly higher disinfection. Conclusions: MPDS containing dual biocides polyquaternium-1 and myristamidopropyl dimethylamine possess the highest disinfection action against multiple ocular pathogens with and without contact lenses when compared with other MPDS. Current single-action polyhexamethylene biguanide solutions are not entirely effective and should not be recommended.