PURPOSE: To report a case of isolated vitritis as the presenting feature of a disseminated infection with Mycobacterium chelonae subspecies gwanakae, a novel subspecies. METHODS: Case report. RESULTS: A 66-year-old male who presented with progressive floaters and blurred vision was found to have isolated vitritis of the left eye. Diagnostic vitrectomy revealed acid-fast bacilli in the vitreous. Whole genome sequencing identified the organism as Mycobacterium chelonae subspecies gwanakae, a recently identified novel subspecies. The patient was admitted for further evaluation and treatment. Punch biopsies of the numerous ulcerative skin lesions revealed acid-fast bacilli. Magnetic resonance imaging revealed osteomyelitis underlying the soft tissue infections, requiring incision and drainage. Biopsy of the bone also revealed acid-fast bacilli. He was treated with weekly intravitreal amikacin and empiric intravenous azithromycin, imipenem, and tobramycin. His ocular infection responded well to treatment with resolution of the vitritis and improvement in visual acuity. Unfortunately, his systemic condition deteriorated, and he declined further intervention, ultimately succumbing to his systemic infection 12 weeks after presentation. CONCLUSION: Isolated vitritis can be the presenting feature of a severe disseminated Mycobacterium chelonae subspecies gwanakae infection.