Xueping Liu

Detection of adenosine using surface-enhanced Raman scattering based on structure-switching signaling aptamer.

In the present study, we report a novel sensitive method for the detection of adenosine using surface-enhanced Raman scattering (SERS) sensing platform based on a structure-switching aptamer. First, Ag-clad Au colloids film on a polished gold disc is prepared as enhanced substrate and modified with thiolated capture DNA. The formation of an aptamer/DNA duplex of expanded anti-adenosine aptamer and tetramethylrhodamine-labeled DNA (denoted TMR-DNA) is then developed, in which TMR-DNA could also hybridize completely with capture DNA. The introduction of adenosine thus triggers structure switching of the aptamer from aptamer/DNA duplex to aptamer/target complex. As a result, the released TMR-DNA is captured onto the SERS substrate, resulting in an increase of SERS signal. Under optimized assay conditions, a wide linear dynamic range (2.0 x 10(-8)M to 2 x 10(-6)M) was reached with low detection limit (1.0 x 10(-8)M). Moreover, high selectivity, stability and facile regeneration are achieved. The successful test demonstrates the feasibility of the strategy for adenosine assay.

Ultrasensitive electrochemical immunosensor for ochratoxin A using gold colloid-mediated hapten immobilization

A convenient, specific, and highly sensitive electrochemical immunosensor based on an indirect competitive assay format was developed for the determination of ochratoxin A (OTA), a common toxic contaminant in various kinds of agricultural products. The sensing substrate was prepared using a gold electrode modified with a self-assembled monolayer of 1,6-hexanedithiol that mediated the assembly of a gold colloid layer, which could enhance the surface loading of OTA-ovalbumin conjugate and improve the sensitivity in electrochemical readouts. After competition of the limited anti-OTA mouse monoclonal antibody between immobilized hapten and OTA analyte in sample solution, alkaline phosphatase (ALP)-labeled horse anti-mouse immunoglobulin G (IgG) antibody was selectively bound onto the surface of the electrode, affording an indicator for OTA concentration in the sample. Electrochemical response arising from the oxidation of enzymatic product of 1-naphthyl phosphate was observed to be inversely proportional to OTA concentration in the range from 10 pg/ml to 100 ng/ml with a detection limit as low as 8.2 pg/ml. Furthermore, a negligible matrix effect and good recoveries were obtained in the determination of corn samples, evidencing the feasibility of the proposed method for accurate determination of OTA in corn samples.

Biocatalyzed deposition amplification for detection of aflatoxin B1 based on quartz crystal microbalance

An ultrasensitive piezoelectric method for the detection of the aflatoxin B1 (AFB1) based on the indirect competitive immunoassay and the biocatalyzed deposition amplification has been developed. In this method, the quartz crystal surface was coated with a self-assembled monolayer of 3-mercaptopropionic acid (MPA) for covalently immobilization of the BSA-AFB1 conjugate, which could compete with the free AFB1 for binding to the anti-AFB1 antibody (MsIgG). After the competitive immunoreaction, the horseradish peroxidase (HRP) labeled goat anti-mouse IgG (G-Anti-MsIgG) was introduced into the detection cell to combine with the anti-AFB1 antibody on the crystal surface. The enzyme labeled G-Anti-MsIgG as a biocatalyst could accelerate the oxidation of 4-chloro-1-naphthol by H2O2 to yield the insoluble product benzo-4-chlorohexadienone on the surface of quartz crystal microbalance (QCM), resulting in a mass increase that was reflected by a decrease in the resonance frequency of the QCM. The proposed approach could allow for the determination of AFB(1) in the concentration range of 0.01-10.0 ng mL(-1). Furthermore, several artificially contaminated milk samples were analyzed with good recoveries obtained, which demonstrated the suitability of the proposed method for detecting AFB1.