A black phosphorus-based system detects micro-RNA in aqueous humor, enabling safe diagnosis of Primary Central Nervous System Lymphoma
Early diagnosis of primary central nervous system lymphoma (PCNSL) remains challenging because brain biopsies are invasive and imaging often lacks molecular specificity. A team led by researchers at Shenzhen University has now developed a minimally invasive fibre-optic plasmonic sensor capable of detecting PCNSL-associated microRNAs in the eye’s aqueous humor with attomolar sensitivity.
At the heart of the approach is a black phosphorus (BP)–engineered surface plasmon resonance (SPR) interface. An ultrathin BP layer is deposited on a gold-coated fiber tip. Because of the work-function difference between BP and gold, electrons transfer from BP into the Au film, creating a strongly enhanced local electric field at the metal–semiconductor interface. This BP–Au charge-transfer nano-interface amplifies refractive-index changes at the surface far more efficiently than conventional metal-only SPR chips, enabling the detection of molecular interactions that would otherwise be too subtle to resolve and pushing the limit of detection down to 21 attomolar without nucleic-acid amplification. The BP layer also provides a high-area, biocompatible surface for immobilizing RNA reporters.
To achieve sequence specificity, the researchers integrated CRISPR-Cas13a, an RNA-guided nuclease that becomes catalytically active only when its target sequence is perfectly matched to a designed CRISPR RNA (crRNA). When the target microRNA (miR-21) is present, activated Cas13a cleaves RNA reporters attached to the BP-modified fiber surface, releasing gold nanoparticles and reducing the local refractive index. The resulting optical shift is read out in real time through the SPR response of the BP-enhanced fiber probe, providing single-nucleotide-resolved detection directly on the plasmonic interface.
With this combined strategy, the sensor achieved a limit of detection of 21 attomolar in buffer and successfully distinguished single-base-mismatched microRNAs. In tests on aqueous-humor samples from patients with PCNSL, the CRISPR-BP-FOSPR assay produced results that closely matched clinical qPCR data, despite operating without any amplification steps.
Because aqueous-humor aspiration is a minimally invasive ophthalmic procedure, this BP-driven plasmonic platform may offer a practical route for early PCNSL screening, longitudinal monitoring, and potentially the diagnosis of other neurological diseases reflected in eye-fluid biomarkers. More broadly, the work showcases how black-phosphorus-based charge-transfer interfaces can be used to engineer next-generation, fibre-integrated biosensors that combine extreme sensitivity with molecular precision.
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