Fungal infection monitoring on corneal epithelium ex vivo model and its collection over polyethersulfone membrane for detecting Candida albicans and Aspergillus fumigatus.

Abstract

In this paper we report a human ex vivo model of Aspergillus fumigatus and Candida albicans infection of the cornea. We demonstrate the ability to monitor fungal growth on the cornea through optical and immunofluorescence microscopy. Additionally, we establish a method for collection of fungal antigens in polyethersulfone (PES) membrane-based sample collectors from the surface of fungal inoculated corneas. Immunofluorescence microscopy was performed directly on both corneal tissue and PES membrane sample collectors. For the latter case, we devised an original ad-hoc method by attaching the membranes used for the collection on standard glass slides, which we call glass slide KIT. This enabled easy handling and improved efficiency for specifically recognizing the corresponding fungal infections by simple immunoassay protocols. As a result, we firstly observe the ability to monitor fungi in corneal tissue, and secondly demonstrate the adsorption of fungal antigens onto PES membranes. Thus, we report for the first time the specific detection of fungal corneal infections in easy to use PES membrane based glass slide KITs through simple immunofluorescence, as an alternative to microbiological culture.