Rapid Diagnosis of Polyfungal Endophthalmitis Utilizing Novel Panfungal and Nested Primers in a Polymerase Chain Reaction (PCR) Assay

Abstract

Abstract: : Purpose: Fungal endophthalmitis is a rare cause of visual loss. The etiologic diagnosis of fungal endophthalmitis has been limited by small specimen size as well as poor sensitivity of currently available diagnostic techniques. The purpose of this study was to apply novel panfungal and nested primers in a PCR assay to assist in the establishment of a rapid and early diagnosis of fungal endophthalmitis. Methods: A 70 year old woman with acute myelogenous leukemia presented with acute bilateral vision loss and vitritis. Bilateral pars plana vitrectomy was performed and vitreous samples were submitted for microbiologic and PCR assays to establish the diagnosis. Novel panfungal and nested primers were used in a PCR assay to identify fungal DNA present in vitreous fluid. Results: Initial gram stains revealed the presence of multiple septated hyphae in vitreous samples from both eyes. Using the panfungal primers, the blood and vitreous were positive for two different fungal DNA bands. These were identified as Fusarium and Candida glabrata using a nested PCR primer assay. Blood and vitreous cultures revealed only the presence of Fusarium species. Conclusions: This assay demonstrates the usefulness of a panfungal and nested primer PCR assay in the establishment of a rapid and early diagnosis of fungal endophthalmitis. The assay is also useful in simultaneously detecting multiple etiologic agents that may be causing a polyfungal endophthalmitis.

Keywords: endophthalmitis • vitreous • fungal disease