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A photo/electrochemical coupling interface of Ru[dcbpy]32+-AMT/Au (AMT; 5-Amino-1,3,4-thiadiazole-2-thiol) was fabricated using a dehydration condensation sulfhydrating method. For the interface functional properties, a combined dual-signal recording (CDSR) method was applied to characterize the response characteristics, and a scanning electrochemical microscopy-electrochemiluminescence (SECM-ECL) imaging was developed to assess the interface distribution uniformity. The interface biosensing compatibility was validated by constructing a simple DNA sensor. The research results show that the interaction between the two functional parameters follows a synergistic effect mechanism in the coupling conditions and an interference effect mechanism in the detection condition. Under optimized conditions, the saturation dual-signal response values are 156.0 and 86.8 µA, respectively. The statistics and imaging comparison analysis validate good interface distribution uniformity and stability performance. The DNA sensor's dual-signal detection limits to the signal probe (SP) are ∼30 fM and 0.3 pM with linear ranges of 100.0 fM âˆ¼ 1.0 nM and 1.0 pM âˆ¼ 10.0 nM, respectively. The fabricated interface exhibits an effective bi-functional response performance compatible with biosensing. The proposed imaging method has a high technical fit for studying photo/electrochemical coupling interfaces and can also provide a reference for other similar coupling interface analyses.

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A study was carried out to investigate the binding mode of aptamer to ampicillin (AMP) and its electrochemical response behavior. The binding mode was confirmed using the molecular dynamics (MD) simulation method to obtain the corresponding binding dynamic change process. Following the confirmed binding mode, a qualitative elucidation was provided on the electrochemical response characteristics of a single-probe aptamer-based folding sensor. The results show that there exist two different binding modes in two different solution systems, Phys2 and H2O (0.1 M NaCl). These two binding modes can respectively induce two different contraction changes, thereby driving the methylene blue (MB)-modified aptamer probe to show a "close-to-interface" convergence behavior with different degrees on the actual electrode surface, which validates two apparently different electrochemical response behavior characteristics of "signal-on" for the sensor. By contrast, H2O (0.1 M NaCl) as the reaction medium is more conducive to the formation of a stable aptamer/AMP complex and the development of a high-sensitivity analytical method with a low detection limit of 0.033 µM. The simulation results effectively support the experimental results, which is helpful in gaining a deeper understanding of the relationship between the signaling mechanism and practical analytical performance for aptamer-based folding sensors at the molecular level.

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By adding 6 thymines to lengthen the parent aptamer combined with the change of "on" and "off" induced by the target for an assistant stem-loop DNA probe (ASP-SLP-MB), a new folding-type electrochemical kanamycin (Kana) aptamer-engineering dual-probe-based sensor (sensor d) was developed. By purposefully reducing the background current and increasing the electron transfer efficiency of methylene blue (MB), the sensor obtained significantly enhanced detection sensitivity compared with non-aptamer-engineering one-probe-based sensor (sensor a). Such efficacy was validated by a big decrease from 530.6 to 210.2 nA for the background current signal and from 360 to 0.3 nM for the detection limit. In addition to the improved sensitivity, the sensor also exhibited good selectivity, anti-fouling detection performance, and potential quantitative analysis ability, showing a feasible potential practical analytical application in real-life complicated samples, for example, milk and serum. The released results prove that the aptamer-engineering method is effective in improving the analytical performance of folding-type sensors and provides a methodological guidance for the design and fabrication of other high-performance folding-type aptasensors. Graphical abstract.

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Two typical kanamycin-A (KAN-A) electrochemical aptamer-based sensors employing different signal transduction mechanisms were deliberately designed and constructed with a similar structure. One sensor (sensor-1) was based on the classical probe conformation changing mode (PCCM) with a methylene blue (MB) label used as an electrochemical tag; the other sensor (sensor-2) used the target-induced signal probe shifting (TISPS) method with a free MB label in the assay solution. The difference in signal transduction mechanisms resulted in big differences in basic electrochemical behavior and comprehensive sensing performance. The results show that both sensor types exhibit different electrochemical behavior in square wave voltammetry, cyclic voltammetry, and in sensitivity, with detection limits of 3.0 nM for sensor-1 and 60.0 pM for sensor-2 in buffer. When validated for practical and quantitative detection of tap water and milk samples, both sensing methods performed well with detection limits of <260 nM and measurement times of <40 min. In addition, accuracy was good with mean recoveries of 72.3-92.6% and precision was acceptable with a relative standard deviation (RSD) of ≤14.2%. The basis for the similarities and differences in performance is also presented.

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Global food and water safety issues have prompted the development of highly sensitive, specific, and fast analytical techniques for food and water analysis. The electrochemical aptamer-based detection platform (E-aptasensor) is one of the more promising detection techniques because of its unique combination of advantages that renders these sensors ideal for detection of a wide range of target analytes. Recent research results have further demonstrated that this technique has potential for real world analysis of food and water contaminants. This review summaries the recently developed E-aptasensors for detection of analytes related to food and water safety, including bacteria, mycotoxins, algal toxins, viruses, drugs, pesticides, and metal ions. Ten different electroanalytical techniques and one opto-electroanalytical technique commonly employed with these sensors are also described. In addition to highlighting several novel sensor designs, this review also describes the strengths, limitations, and current challenges this technology faces, and future development trend.

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