This case involved a 2-year-old neutered male domestic mixed-breed cat that was rescued from the street eight months earlier. The animal presented with weakness, hyporexia, progressive weight loss, fatigue, uveitis, pale mucous membranes, dehydration (7%), and pelvic limb paresis. Aqueous humor was collected for molecular analysis for the differential diagnosis of potential etiological agents [Feline coronavirus (FCoV), Feline leukemia virus (FeLV), Feline immunodeficiency virus (FIV), Toxoplasma gondii, Cryptococcus spp., Felid herpesvirus-1 (FHV-1) and Bartonella spp.] of feline uveitis. The sample was positive by real-time reverse transcription-polymerase chain reaction (RT-qPCR) for FCoV and RT-qPCR and real-time polymerase chain reaction (qPCR) for FeLV and qPCR FIV. The cat was euthanized due to poor clinical outcomes and prognosis. A cerebrospinal fluid (CSF) sample was collected and tested, and the same pathogens were found in the aqueous humor. Small-cell follicular multicenter lymphoma and multifocal pyogranulomatous meningoencephalitis were observed upon histopathological analysis. In this study, aqueous humor and cerebrospinal fluid samples were efficient for the detection of coinfection with FIV, FeLV, and FCoV.