Portable multichannel detection instrument based on time-resolved fluorescence immunochromatographic test strip for on-site detecting pesticide residues in vegetables.

In this work, a portable multichannel detection instrument based on time-resolved fluorescence immunochromatographic test strip (TRFIS) was proposed for on-site detecting pesticide residues in vegetables. Its hardware consisted of a silicon photodiode and excitation light source array, a mainboard of the lower machine with STMicroelectronics 32 (STM32) and a linear stepping motor. While detecting, cardboard with 6-channel TRFIS was pulled into the cassette by the stepping motor. The peak area of the test (T) line and control (C) line of each TRFIS was sampled and calculated by software, then the concentration of the detected pesticide was obtained according to the ratio of the T to C value. This instrument could sample 6-channel TRFIS within 30 s simultaneously, and it exhibited excellent accuracy with a 2.5% average coefficient of variation for each channel (n = 12). In addition, the TRFIS was constructed by using europium oxide time-resolved fluorescent microspheres to label the monoclonal antibody against acetamiprid and form a fluorescent probe, which was fixed on the binding pad. The TRFIS was used for the detection of acetamiprid in celery cabbage, cauliflower and baby cabbage. This instrument was used to complete the qualitative and quantitative analysis of the TRFIS, so as to enhance the practical application of the detection method. This TRFIS possessed excellent linearity ranging from 0.25 mg kg-1 to 1.75 mg kg-1 for the detection of acetamiprid, and the limit of detection were 0.056-0.074 mg kg-1 in the different vegetable matrix. The platform combines the accuracy and portability of traditional test strips with the highly sensitive and efficient fluorescence intensity recognition function of detection equipment, which shows a great application prospect of multi-channel rapid detection of small molecule pollutants in the field.

Portable electrochemiluminescence detection system based on silicon photomultiplier single photon detector and aptasensor for the detection of tetracycline in milk.

In this work, a portable electrochemiluminescence (ECL) detection system based on silicon photomultiplier (SiPM) single photon detector was proposed for the detection of ECL signals on a screen-printed electrode (SPE). This instrument innovatively used SiPM single photon detector to detect the ECL signal, which solved friability and bloat caused by the high operating voltage and the limitation of detection components in the traditional ECL detection instrument. This detection instrument showed excellent electrochemical and ECL detection performance. On this basis, an aptasensor based on silver (core)-gold (shell) bimetallic nanoparticles (Ag@AuNPs) was constructed for the detection of tetracycline (TET) in milk on SPE. Here, Ag@AuNPs had a significant effect in enhancing luminol ECL signal and immobilizing aptamer. The concentration of TET was detected according to the changes of the ECL signal intensity of the detection instrument. This instrument exhibited an excellent linearity ranging from 0.01 ng/mL to 1,000 ng/mL for the detection of TET, and a limit of detection (LOD) was 0.0053 ng/mL. The developed portable ECL detection instrument provides a new platform for the detection of small molecule contaminants.

A novel label-free electrochemiluminescence aptasensor using a tetrahedral DNA nanostructure as a scaffold for ultrasensitive detection of organophosphorus pesticides in a luminol-H2O2 system.

In this work, a new type of Au-tetrahedral DNA nanostructure (Au-TDN) was originally proposed and successfully applied in an electrochemiluminescence aptasensor to detect organophosphorus pesticides (Ops). The aptamers modified with -SH could be covalently bonded with gold nanoparticles (AuNPs) to form a tetrahedron structure, and there were independent probes at each vertex of the tetrahedron, which could increase the probability of specific binding with Ops. The originally designed structure could not only maintain a stable tetrahedral configuration, but also combined with the target to improve the sensitivity of the sensor. Meanwhile, silver nanoparticles (AgNPs) could catalyze the chemical reaction between luminol and H2O2 to generate a variety of intermediates called reactive oxygen species (ROS) for signal enhancement. Factors that had important influences on the aptasensor, such as the concentration of Au-TDN, the incubation time, and the pH value of the buffer, were optimized in this trial. According to the final results, the limit of detection (LOD) of 3 pg mL-1 (S/N = 3) for methyl parathion, the LOD of 0.3 pg mL-1 (S/N = 3) for parathion and the LOD of 0.03 pg mL-1 (S/N = 3) for phoxim were obtained, respectively. Moreover, the novel tetrahedral structure could be replaced by different types of aptamers to expand its application range and lay a foundation for the development of portable rapid detection devices for pesticide residues.

A novel electrochemiluminescence aptasensor based on copper-gold bimetallic nanoparticles and its applications.

In this work, a novel electrochemiluminescence (ECL) aptasensor was structured for the detection of four organophosphorus pesticides (OPs). Firstly, multi-walled carbon nanotubes (MWCNTs) were used to create a favorable loading interface for the fixation of tris (2, 2'-bipyridyl) ruthenium (II) (Ru (bpy)32+). At the same time, copper (core)-gold (shell) bimetallic nanoparticles (Cu@Au NPs) were synthesized in the aqueous phase for the sensor construction. Gold nanoparticles (Au NPs) could promote the electrochemiluminescence intensity of Ru (bpy)32+ with high efficiency by catalyzing the oxidation process of tri-n-propylamine (TPrA). Compared with the Au NPs, Cu@Au NPs increased the solid loading of Au NPs by virtue of the large specific surface area of copper nanoparticles (Cu NPs), which could further improve the sensitivity of aptasensor. When OPs were added, the ECL intensity was significantly reduced, and the concentration of OPs could be detected through the ECL intensity. Under the optimum conditions, the aptasensor had a wider dynamic range and ultra-low detection limit for the detection of four pesticides: profenofos, isocarbophos, phorate, and omethoate, and their detection limits were 3 × 10-4 ng/mL, 3 × 10-4 ng/mL, 3 × 10-3 ng/mL, and 3 × 10-2 ng/mL respectively (S/N = 3). The aptasensor had the merits of good stability, reproducibility, and specificity, and had a favorable recovery rate in detecting OPs residues in vegetables. This work provided an effective method for the construction of a simple, rapid, and sensitive biosensor.

Novel Au-tetrahedral aptamer nanostructure for the electrochemiluminescence detection of acetamiprid.

In this work, an ultrasensitive and selective electrochemiluminescence (ECL) aptasensor with Au-tetrahedral aptamer nanostructure (Au-TAN) for acetamiprid detection was developed, which employed luminescence property of luminol and hydrogen peroxide (H2O2) as a co-reactant to apply the prepared Au-TAN to the luminescence systems. Au-TAN was prepared to modify an electrode surface via an Au-S bond to form a stable tetrahedral nanostructure. Fixed on the surface of the working electrode, Au-TAN could not only enhance the function of the aptamer but also boost the sensing performance. At the same time, Au nanoparticles (AuNPs) of the Au-TAN could also catalyze H2O2, thereby enhancing the luminescence performance of this aptasensor. The pH of the buffer solution, the concentration of H2O2 and the concentration of Au-TAN were optimized. Under the optimal conditions, the aptasensor had a detection limit of 0.0576 pM (S/N = 3), which was lower than those of other aptasensors for acetamiprid detection. Moreover, the weak alkaline environment explored in the experiment could expand its application range. Above all, the proposed method presented a high accuracy and sensitivity.

Broad-spectrum electrochemical immunosensor based on one-step electrodeposition of AuNP-Abs and Prussian blue nanocomposite for organophosphorus pesticide detection.

Broad-spectrum antibodies can effectively recognize substances with similar structures and have broad application prospects in field rapid detection. In this study, broad-spectrum antibodies (Abs) against organophosphorus pesticides (OPs) were used as sensitive recognition elements, which could effectively recognize most OPs. Gold nanoparticles (AuNPs) have good biocompatibility. It combined with Abs to form a gold-labeled probe (AuNPs-Abs), which enhances the effective binding of antibodies to nanomaterials. Prussian blue (PB) was added to electrodeposition solution to enhance the conductivity, resulting in superior electrochemical performance. The AuNP-Abs-PB composite film was prepared by electrodeposition on the electrode surface to improve the anti-interference ability and stability of the immunosensor. Under the optimal experimental conditions, the immunosensor had a wide detection range (IC20-IC80: 1.82 × 10-3-3.29 × 104 ng/mL) and high sensitivity. Most importantly, it was simple to be prepared and could be used to detect multiple OPs.

Novel Time-Resolved Fluorescence Immunochromatography Paper-Based Sensor with Signal Amplification Strategy for Detection of Deoxynivalenol.

Immunoassay has the advantages of high sensitivity, high specificity, and simple operation, and has been widely used in the detection of mycotoxins. For several years, time-resolved fluorescence immunochromatography (TRFIA) paper-based sensors have attracted much attention as a simple and low-cost field detection technology. However, a traditional TRFIA paper-based sensor is based on antibody labeling, which cannot easily meet the current detection requirements. A second antibody labeling method was used to amplify the fluorescence signal and improve the detection sensitivity. Polystyrene fluorescent microspheres were combined with sheep anti-mouse IgG to prepare fluorescent probes (Eu-IgGs). After the probe fully reacted with the antibody (Eu-IgGs-Abs) in the sample cell, it was deployed on the paper-based sensor using chromatography. Eu-IgGs-Abs that were not bound to the target were captured on the T-line, while those that were bound were captured on the C-line. The paper-based sensor reflected the corresponding fluorescence intensity change. Because a single molecule of the deoxynivalenol antibody could bind to multiple Eu-IgGs, this method could amplify the fluorescence signal intensity on the unit antibody and improve the detection sensitivity. The working standard curve of the sensor was established under the optimum working conditions. It showed the lower limit of detection and higher recovery rate when it was applied to actual samples and compared with other methods. This sensor has the advantages of high sensitivity, good accuracy, and good specificity, saving the amount of antibody consumed and being suitable for rapid field detection of deoxynivalenol.