Background-Free Chromatographic Detection of Sepsis Biomarker in Clinical Human Serum through Near-Infrared to Near-Infrared Upconversion Immunolabeling

Luminescence nanomaterial-based lateral flow assay (LFA) is promising for point-of-care tests. However, the detection sensitivity and accuracy are often affected by the interferences of autofluorescence and photon scattering from nitrocellulose membrane and colored plasma. Here, we describe a near-infrared to near-infrared upconversion nanoparticle (UCNP) immunolabeled LFA for background-free chromatographic detection of sepsis biomarker procalcitonin (PCT) in clinical human plasma. This upconversion immunolabeling enables both light excitation (at ∼980 nm) and anti-Stokes emission (at 800 nm) to be adopted within the first biological window (700–1000 nm), which eliminates background autofluorescence as well as photon scattering interferences, empowering a high-sensitivity detection without complicated procedures. After optimization, the described assay presented a limit of detection reaching down to 0.03 ng/mL, lower than the normal level (0.05 ng/mL), while having a detection range of 0.03–50 ng/mL that covers the clinical PCT level of interest (0.5–10 ng/mL). The assay recoveries in human serum samples were evaluated to be about 95–110%, whereas the inter- and intra-assay coefficient variations were both determined to be below 15%. Importantly, measured PCT concentrations in clinical samples are in good correlation with that of the electrochemiluminescence immunoassay (Roche) widely applied in large clinical settings. This near-infrared to near-infrared upconversion immunolabeling approach has direct implications for ultrasensitive and background-free point-of-care detection of other serum biomarkers in resource-limited clinical settings.