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Antifungal Susceptibility Test via Single-Cell Morphology, Development, and Metabolism

Xiao Han,Yanmei Zhang,Xiaoshan Zheng,Xixian Wang,Rongze Chen,Min Liu,Qiwen Yang,Guanghua Huang,Yu Vincent Fu,Bo Ma,Jiadong Huang,Jian Xu

Antifungal susceptibility test (AFST) is essential for guiding effective therapy, but it remains challenged by long turnaround times and the limited generalizability of single-parameter phenotypic readouts across diverse fungal species and antifungal classes. Here, we present a multifeature AFST (MAFST) platform that integrates single-cell Raman spectroscopy and brightfield imaging to enable rapid and pleiotropic assessment of antifungal responses. By unifying complementary metabolic and morphological features into a composite inhibition index (CII), MAFST captures coordinated cellular responses to antifungal exposure that cannot be reliably resolved by individual metrics alone. Benchmarking against the broth microdilution demonstrates that CII-based MAFST achieves high concordance across multiple Candida species and antifungal classes while substantially reducing the time required for susceptibility determination. Thus, by capturing single-cell morphological, developmental, and metabolic drug responses in a fast, accurate, and widely applicable fashion, MAFST can contribute to tackling fungal infections.