An electrochemical paper-based hydrogel immunosensor to monitor serum cytokine for predicting the severity of COVID-19 patients.

Analysis of cytokines levels in human serum is critical as it can be a "symptom diagnostic biomarker" in COVID-19, giving real-time information about human health status. Here, we present the construction and performance of a low-price immunosensor (∼US$0.428 per test) based on microfluidic paper-based system to detect cytokine for predicting the health status of COVID-19 patients. Interleukin-6 (IL-6) was selected as the detection model for the close relationship between IL-6 and COVID-19. The assay, which we integrated into foldable paper system, leverages the magnetic immunoassay, the streptavidin-horseradish peroxidase (HRP) associated with tetramethyl benzidine/hydrogen peroxide (TMB/H2O2) to amplify the signal for electrochemical readout. To improve the sensitivity of cytokine detection, a hybrid of gold nanoparticles (AuNPs) and polypyrrole (PPy) hydrogel was modified on the working electrode to increase the conductivity and improve the electron transfer rate. With our prototypic origami paper-based immunosensor operated in differential pulse voltammetry (DPV) mode, we achieved excellent results with a dynamic range from 5 to 1000 pg/mL and a lower detection limit (LOD) of 0.654 pg/mL. Furthermore, we evaluated the capability of the clinical application of the proposed immunosensor using human serum samples from a hospital. The results indicate that our proposed immunosensor has great potential in early diagnosing high-risk COVID-19 patients.

Microfluidic electrochemical magnetoimmunosensor for ultrasensitive detection of interleukin-6 based on hybrid of AuNPs and graphene.

Cytokines are important factors in the early diagnosis of autoimmune diseases and require high sensitivity, high selectivity and quantitative detection. We proposed a miniaturized electrochemical magneto-immunosensor (EC-MIS) on portable interleukin-6 (IL-6) detection based on this requirement. Firstly, a micro-fabricated working electrode is electrochemically modified with a hybrid of reduced graphene oxide (rGO) and gold nanoparticles (AuNPs). Increased surface area and enhanced charge transfer rate improve the performance of this immunosensor on sensitivity. Secondly, magnetic beads attached with the capture antibody (cAb) are employed in sandwich immunoassay. This kind of immunoassay is immobilized on the working electrode surface by an external magnet to enrich the analyte IL-6. Thirdly, the last two features are combined and integrated on a microfluidic device in order to restrict the sample at certain areas and ease the operation of detection. With our prototypic EC-MIS operated in amperometric mode, we have achieved the detection of IL-6 with a linear range from 0.97 to 250 pg/mL and a limit of detection (LOD) of 0.42 pg/mL. Real serum samples were demonstrated and compared with benchtop equipment's results.