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It is known that gold nanoparticles (AuNPs) possess peroxidase-like activity. They can catalyze the oxidation of 3,3,5,5-tetramethylbenzidine by HO which leads to a color change from red to blue. It is shown here that the peroxidase-like activity of AuNPs can be inhibited by passivating its surface passivation with a ssDNA aptamer against sulfadimethoxine. If, however, the target molecule (sulfadimethoxine) is present, the aptamer is desorbed from the AuNPs surface, and this results in the reactivation of the catalytic property of the AuNPs. The color change of the solution (from purple to blue) is related to the analyte concentration, and this can be judged visually or by UV-visible absorptiometry at 650 nm. The assay, under optimized conditions, has a detection limit of 10 ng·mL of sulfadimethoxine, and the calibration plot is linear over a rather wide concentration range (0.01-1000 μg·mL). The assay can be performed within <15 min, is sensitive, and therefore is well suited for fast screening in food analysis. Conceivably, it can be extended to many other small analytes for which aptamers are available. [ABSTRACT FROM AUTHOR]

Most aptasensors are designed based on aptamers' conformational change or structural transformation upon interaction with their targets. A limitation of these strategies is the unpredictability of such structural alterations that may yield false-positive or nonspecific signals. In bioanalysis, a dominant strategy for protein determination is the sandwich assay because of its high specificity and sensitivity resulting from the dual recognition mechanism. However, the approach is poorly suited to detection of low molecular weight targets, which unlikely bind to two aptamers simultaneously due to steric hindrance. To circumvent this drawback, a split aptamer strategy was recently developed, in which nucleic acid aptamers were split into two fragments that could specifically form a ternary assembly in the presence of ligand. This elegant approach has been extensively adopted for detection of various targets, especially small molecular weight targets, with different transduction methods including colorimetric, fluorescence, and electrochemical techniques. This review first introduced split aptamers and how to obtain split aptamers, and then summarized recent advances in the development of sandwich-format biosensing assays based on split aptamer fragments, aiming at providing a general guide for the design of split-aptamer-based sandwich assays. Furthermore, we discussed possible challenges in the development of split-aptamer-based sandwich assay regarding stability, sensitivity and multiplicity, so as to offer future opportunities of this assay. [ABSTRACT FROM AUTHOR]

Aptamer-based strip assay is an easy, highly efficient and low-cost detection method, which has been developed and easily applied to onsite detection. A new sensitive sandwich dipstick assay for adenosine triphosphate (ATP) detection was successfully developed based on specific recognition between split aptamer fragments and the target. In this method, the thiolated aptamer was first conjugated to the surface of gold nanoparticles (AuNPs), while the biotin-aptamer was immobilized on the surface of a nitrocellulose filter in the test line. In the presence of ATP, the thiol-aptamer/ATP/biotin-aptamer complexes were generated, which led to an obvious increase in optical signals at the test line. Under the optimal determination conditions, an excellent linear logarithmic response to the ATP concentration was obtained within the range of 0.5 μM to 5 mM. The limit of detection (LOD) of 0.5 μM was reached at a signal-to-noise ratio of 3. The dipstick assay showed a good average recovery of 96-108 % with the RSD of less than 20 % in urine samples. The proposed method exhibited high specificity against other nucleotides such as the uridine triphosphate (UTP), cytidine triphosphate (CTP), and guanosine triphosphate (GTP). The results indicated that the dipstick strip may be considered as an inexpensive screening tool for onsite ATP determination. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]