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

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.

抗真菌药物敏感性试验（Antifungal Susceptibility Test, AFST）对于指导临床抗真菌精准治疗至关重要，但该方法仍面临周转周期漫长、单参数表型读数在不同真菌物种与抗真菌药物类别间泛化能力有限等挑战。本研究提出一种集成单细胞拉曼光谱与明场成像的多参数抗真菌药物敏感性试验（MAFST）平台，可对抗真菌药物应答进行快速且多维度的综合评估。该平台将互补的代谢与形态学特征整合为综合抑制指数（CII），能够捕捉到单一指标无法可靠解析的抗真菌药物暴露下的协同细胞应答。以肉汤微量稀释法作为金标准进行基准测试的结果显示，基于CII的MAFST在多种念珠菌属菌株与不同抗真菌药物类别中均表现出极高的一致性，同时大幅缩短了药敏检测所需的时间。因此，MAFST能够以快速、准确且普适性强的方式获取单细胞层面的形态、发育与代谢药物应答特征，可为真菌感染防控工作提供有力支撑。