Simultaneous Quantitative Detection of IL-6 and PCT Using SERS magnetic immunoassay with sandwich structure

Sepsis is a systemic inflammatory response syndrome caused by infection. The mortality rate is as high as 30%-50%. Early diagnosis and treatment can significantly improve the mortality of patients with sepsis. Therefore, we have developed a SERS-based magnetic immunoassay method that uses the principle of sandwich method to quantitatively detect Interleukin 6 (IL-6) and Procalcitonin (PCT). In this article, two different Raman reporter molecules are embedded in the middle of the Au@Ag shell and coupled with the tracer antibody to form a SERS immunoprobe. Biotin was coupled with capture antibody to form a sandwich structure when participating in the immune response. Streptavidin and biotin systems have extremely high binding affinity. The sandwich structure is quickly captured by SA magnetic beads and then applied with a magnetic field to enrich the magnetic beads. Finally, simultaneous quantitative detection is achieved by the intensity of the two Raman reporter characteristic peaks on the solution magnetic beads. IL-6 and PCT showed a good relationship between 0-1000 pg ml^-1and 0-20 ng ml^-1, respectively, and the limits of detection were 0.54 pg ml^-1and 0.042 ng ml^-1, respectively. The recovery rate was between 89.8% and 104.2%, both intra-assay and inter-assay CV were ≤20%. No cross-reaction with C-reactive protein (100μg ml^-1), showing good specificity. This method provides a new technical reference for the clinical detection of sepsis biomarkers.