Ovarian cancer remains the deadliest gynecological malignancy, largely because early screening is inadequate. This work reports a smartphone-based fluorescent homogeneous immunoassay sensor (SFHIS) that combines aggregation-induced emission luminogen-doped metal-organic framework microspheres (AMOFs) as donors and spiky carbon nanospheres (CNPs) as acceptors. Target antigen–antibody sandwich formation brings donor and acceptor into proximity, inducing efficient Förster resonance energy transfer (FRET) and fluorescence quenching, which is linearly related to the natural logarithm of antigen concentration. A dual-correction algorithm removes internal filter effects, enabling accurate quantification. The SFHIS shows linear ranges of 0.8−51.2 U/mL for CA125 and 8–256 pmol/L for HE4, with limits of detection of 0.67 U/mL and 1.03 pmol/L, respectively. The assay is completed within 25 min without washing, exhibits intra- and inter-batch CVs < 10%, no cross-reactivity with CEA, AFP, or PSA, and correlates strongly with chemiluminescence assays (CA125: R2 = 0.970; HE4: R2 = 0.982; P < 0.05). This portable, low-cost platform offers rapid, reliable biomarker detection suitable for clinical laboratories, primary care, community health, and resource-limited settings.
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