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C-reactive protein (CRP) is a widely accepted biomarker of cardiovascular disease and inflammation. In this study, a RNA aptamer-based electrochemical sandwich type aptasensor for CRP detection was described using the functionalized silica microspheres as immunoprobes. Silica microspheres (Si MSs), which have good monodispersity and uniform shape, were firstly synthesized. The silica microspheres functionlized with gold nanoparticles (Au NPs) provided large surface area for immobilizing signal molecules (Zinc ions, Zn 2+ ) and antibodies (Ab). RNA aptamers, which were specific recognized to CRP, were assembled on the surface of Au NPs modified electrode via gold-sulfur affinity. In the presence of CRP, a sandwich structure of aptamer-CRP-immunoprobe was formed. Square wave voltammetry (SWV) was employed to record the sensing signal, and a clearly reductive peak corresponding to Zn 2+ at about -1.16V (vs. SCE) was obtained. Under optimal conditions, the aptasensor showed wide linear range (0.005ngmL -1 to 125ngmL -1 ) and low detection limit (0.0017ngmL -1 at a signal-to-noise ratio of 3). Some possible interfering substance was also investigated, and the results obtained showed that the aptasensor possessed good selectivity. When the aptasensor was applied to real serum samples analysis, the satisfied results were obtained, indicating that the aptasensor possessed potential real application ability., (Copyright © 2017 Elsevier B.V. All rights reserved.)

In this study, we tried coupling the small signal molecules that could intercalate into DNA double helix with hybridization chain reaction (HCR) technique to fabricate a multiple immunosensor. Doxorubicin hydrochloride (DXH) and methylene blue (MB) were used as signal molecules and alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) were selected as model biomarkers. The immunosensor mainly consists of three parts as follows: First, two different primary antibodies (Ab 1 ) immobilized on the surface of gold nanoparticles (Au NPs); Second, secondary antibodies (Ab 2 ) conjugated with DNA primer; Third, long DNA concatemers from HCR were used as a carrier to intercalate amounts of signal molecules (DXH or MB). A sandwich immunocomplex was formed among primary antibodies, target biomarkers and secondary antibodies conjugated with DNA primer via specific recognition reaction. Afterwards, DNA concatemers intercalating amounts of DXH or MB were linked to DNA primer via DNA hybridization. Square wave voltammetry (SWV) was employed to record the response signals from electroactive molecules DXH and MB, and two distinguishable signals were obtained, which peak potentials were at about -0.30V (corresponding to MB) and -0.70V (corresponding to DXH, both vs SCE), respectively. The signal intensities of MB and DXH were linearly related to the logarithm of biomarkers concentration in the range of 0.05pgmL -1 -25ngmL -1 , and the limit of detection were 0.03pgmL -1 for CEA and 0.02pgmL -1 for AFP (at S/N=3), respectively. Furthermore, the immunosensor exhibited a sensitive electrochemical response to biomarkers in human serum samples and the results obtained were in accordance with reference method, indicating the immunosensor can be applied to real sample analysis in clinic diagnosis., (Copyright © 2016 Elsevier B.V. All rights reserved.)