CRISPR-Cas12a immunosensing on glass fiber for point-of-care quantification of multiple inflammation biomarkers in osteoarthritis
Siyi Zou, Jingnan Li, Sheng Lu, Danyu Li, Yuxuan Liu, Weiguo Zhang, Xiaolin Cui, J. Justin Gooding, Kang Tian, Guozhen Liu
Abstract
Inflammation quantification through the detection of cytokines and inflammatory factors in a point-of-care scenario is crucial for managing and preventing diseases. Achieving early-stage inflammation quantification in osteoarthritis (OA), before the radiographic findings become apparent, may provide an opportunity for accurate diagnosis and effective therapy, potentially preventing joint degeneration. Here, we developed a glass-fiber-based universal system with CRISPR-Cas12a-based immunosensors (GUSCI) for simultaneous detection of multiple inflammation biomarkers using a portable fluorescence detection device. GUSCI enables the detection of five inflammation biomarkers (interleukin-4 [IL-4], IL-6, tumor necrosis factor alpha [TNF-α], interferon γ [IFN-γ], and matrix metalloproteinase 3 [MMP-3]) with picomolar sensitivity in a 10-μL sample while having less than 75 min of assay time and a smartphone signal readout. A cohort study of 35 synovial samples from OA patients revealed differences in inflammation levels. This "sample in, answer out" platform not only aids in understanding OA progression but also offers a versatile strategy for point-of-care diagnosis of a wide range of diseases in resource-limited settings.