Silicon-Based Optical Sensors for Fungal Pathogen Diagnostics

Abstract

The last years have witnessed a link between the COVID-19 pandemic with increasing numbers of vulnerable patients and globally emerging incidences of severe drug-resistant fungal infections, thus, calling for rapid, reliable, and sensitive diagnostic tools for fungal infections. However, despite strong warnings from health authorities, such as the World Health Organization, concerning the fatal consequences of the global spread of drug-resistant pathogenic fungi, progress in fungal infection diagnosis and therapy is still limited. Today, gold standard methods for revealing resistance and susceptibility in pathogenic fungi, namely antifungal susceptibility testing (AFST), require several days for completion, and thus this lengthy process can adversely affect antifungal therapy and further promote the spread of resistance. In this work, the use of photonic silicon chips consisting of micropatterned diffraction gratings as sensitive sensors for rapid AFST of clinically relevant fungal pathogens is investigated. These photonic chips provide a surface for the colonization of microbial pathogens at a liquid-solid interface and serve as the optical transducer element for label-free monitoring of fungal growth by detecting real-time changes in the white light reflectance. These sensor elements are used to track morphological changes of fungi in the presence of clinically relevant antifungals at varying concentrations to rapidly determine the minimum inhibitory concentration (MIC) values that help to classify pathogens as resistant or susceptible. We show that by careful design of the chip dimensions, this optical method can extend from bacteria, through yeasts, to filamentous fungi for accelerated AFST, which is at least three times faster than current gold standard methods and can provide same-day results. Moreover, a 3D-printed microfluidic gradient generator was designed to complement the assay and provide an integrated system, which can potentially be employed in point-of-care settings. This gradient generator produces the two-fold dilution series of clinically relevant antimicrobials in an automated manner and is interfaced with the photonic silicon chips to include a complete, on-chip, label-free, and phenotypic assay. Using the bacterial species Escherichia coli and ciprofloxacin as a model pathogen-drug combination, MIC values can be expeditiously determined within 90 minutes compared to current clinical practices, which typically require up to 24 h for bacterial species.