Insight into a Target-Induced Photocurrent-Polarity-Switching Photoelectrochemical Immunoassay for Ultrasensitive Detection of Escherichia coli O157:H7.

Sensitive, portable methods of detection for foodborne pathogens hold great significance for the early warning and prevention of foodborne diseases and environmental pollution. Restricted by a complicated matrix and limited signaling strategies, developing a ready-to-use sensing platform with ultrahigh sensitivity remains challenging. In this work, near-infrared (NIR) light-responsive AgBiS2 nanoflowers (NFs) and Cu2O nanocubes (NCs) were introduced to construct a novel target-induced photocurrent-polarity-switchable system and verified for the development of an all-in-one, ready-to-use photoelectrochemical (PEC) immunosensor. NIR-responsive n-type AgBiS2 NFs and p-type Cu2O NCs producing anodic and cathodic photocurrents were conjugated with monoclonal (MAb1) and polyclonal antibodies (PAb2), respectively. Using a sandwich-type immunocomplex bridged by Escherichia coli O157:H7, an efficient photocurrent-polarity-switching PEC system was formed on a paper-based working electrode (PWE). Owing to the spatial separation of the photogenerated carriers and the elimination of false-positive/negative signals by the polarity-switchable photocurrent, the proposed NIR PEC immunoassay for E. coli O157:H7 exhibits a considerably low detection limit of 8 colony-forming units/milliliter (CFU/mL) with a linear range from 25 to 5 × 107 CFU/mL. The platform includes a PWE with an automatic cleaning function and a portable PEC analyzer with smartphone-compatible Bluetooth capability, thus achieving point-of-care testing of E. coli O157:H7. The sensor was applied to the analysis of pork samples artificially contaminated with E. coli O157:H7, and the detection results were in good agreement with the plate counting method, a gold standard in the field. This work aimed to investigate the photoelectric activity of the NIR-responsive p/n-type composites and to provide a new signal-reversal route for the construction of an all-in-one ready-to-use PEC immunosensor for the detection of low-concentration biomolecules.

A Collection of RPA-Based Photoelectrochemical Assays for the Portable Detection of Multiple Pathogens.

Portable, ultrasensitive, and simultaneously quantitative detection of the nucleic acids of multiple foodborne pathogens is critical to public health. However, the current testing methods depend on costly equipment and tedious amplification steps. In this study, we propose a photoelectrochemical (PEC) biosensor combined with recombinase polymerase amplification (RPA) technology (RPA-PEC) for the rapid detection of multiple foodborne pathogens under irradiation of 980 nm light. In particular, two working surfaces were designed on homemade three-dimensional screen-printed paper-based electrodes. The genomic DNAs of Escherichia coli O157:H7 and Staphylococcus aureus was initiated by RPA on the corresponding electrode surfaces, thus forming a lab-on-paper platform. Using the formed DNA-PEC signaler, photocurrents were achieved at 37 °C after only 20 min of RPA. The detection performance was superior to that of conventional agarose gel electrophoresis, with detection limits of 3.0 and 7.0 copies/µL for E. coli O157:H7 and S. aureus, respectively. Our study pioneers a new RPA-PEC method for foodborne pathogens and provides directions for the construction of lab-on-paper platforms for the portable detection of multiple nucleic acids.

ZnO@Ag-Functionalized Paper-Based Microarray Chip for SERS Detection of Bacteria and Antibacterial and Photocatalytic Inactivation.

Bacterial infections caused by pathogenic microorganisms have become a serious, widespread health concern. Thus, it is essential and required to develop a multifunctional platform that can rapidly and accurately determine bacteria and effectively inhibit or inactivate pathogens. Herein, a microarray SERS chip was successfully synthesized using novel metal/semiconductor composites (ZnO@Ag)-ZnO nanoflowers (ZnO NFs) decorated with Ag nanoparticles (Ag NPs) arrayed on a paper-based chip as a supporting substrate for in situ monitoring and photocatalytic inactivation of pathogenic bacteria. Typical Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli and Vibrio parahemolyticus were selected as models. Partial least-squares discriminant analysis (PLS-DA) was performed to minimize the dimensionality of SERS spectra data sets and to develop a cost-effective identification model. The classification accuracy was 100, 97.2, and 100% for S. aureus, E. coli, and V. parahemolyticus, respectively. The antimicrobial activity of ZnO@Ag was proved by the microbroth dilution method, and the minimum inhibitory concentrations (MICs) of S. aureus, E. coli, and V. parahemolyticus were 40, 50, and 55 µg/mL, respectively. Meanwhile, it demonstrated remarkable photocatalytic performance under natural sunlight for the inactivation of pathogenic bacteria, and the inactivation rates for S. aureus, E. coli, and V. parahemolyticus were 100, 97.03 and 97.56%, respectively. As a result, the microarray chip not only detected the bacteria with high sensitivity but also confirmed the antibacterial and photocatalytic sterilization properties. Consequently, it offers highly prospective strategies for foodborne diseases caused by pathogenic bacteria.

Advances in surface-enhanced Raman spectroscopy technology for detection of foodborne pathogens.

Rapid control and prevention of diseases caused by foodborne pathogens is one of the existing food safety regulatory issues faced by various countries and has received wide attention from all sectors of society. The development of rapid and reliable detection methods for foodborne pathogens remains a hot research area for food safety and public health because of the limitations of complex steps, time-consuming, low sensitivity, or poor selectivity of commonly used methods. Surface-enhanced Raman spectroscopy (SERS), as a novel spectroscopic technique, has the advantages of high sensitivity, selectivity, rapid and nondestructive detection and has exhibited broad application prospects in the determination of pathogenic bacteria. In this study, the enhancement mechanisms of SERS are briefly introduced, then the characteristics and properties of liquid-phase, rigid solid-phase, and flexible solid-phase are categorized. Furthermore, a comprehensive review of the advances in label-free or label-based SERS strategies and SERS-compatible techniques for the detection of foodborne pathogens is provided, and the advantages and disadvantages of these methods are reviewed. Finally, the current challenges of SERS technology applied in practical applications are listed, and the possible development trends of SERS in the field of foodborne pathogens detection in the future are discussed.

SERS Sensors Based on Aptamer-Gated Mesoporous Silica Nanoparticles for Quantitative Detection of Staphylococcus aureus with Signal Molecular Release.

This work describes a simple and novel biosensor for the quantitative determination of Staphylococcus aureus (S. aureus) based on target-induced release of signal molecules from aptamer-gated aminated mesoporous silica nanoparticles (MSNs) coupled with surface-enhanced Raman scattering (SERS) technology. MSNs were synthesized and then modified with amino groups by (3-aminopropyl) triethoxysilane to make them positively charged. Next, signal molecules (4-aminothiophenol, 4-ATP) were loaded into the pores of MSNs. Then, negatively charged aptamers of S. aureus were assembled on the surface of MSNs through electrostatic interactions. Upon the addition of S. aureus, the assembled aptamers were specifically bound to the bacteria. Consequently, the "gates" were opened, resulting in the release of 4-ATP from the pores of MSNs. The released molecules were measured by a Raman spectrometer, and the intensity of 4-ATP at 1071 cm-1 was linearly related to the S. aureus concentration. A silver nanoflower silica core-shell structure (Ag NFs@SiO2) was prepared and it served as a SERS substrate. Under optimized experimental conditions, a good linear relationship (y = 2107.93 + 1536.30x, R2 = 0.9956) in the range from 4.7 × 10 to 4.7 × 108 cfu/mL was observed with a limit of detection of 17 cfu/mL. The method was successfully applied for the analysis of S. aureus in fish samples and the recovery rate was 91.3-109%.

Generalized ratiometric surface-enhanced Raman scattering biosensor for okadaic acid in food based on Au-triggered signal amplification.

BACKGROUND: Reliability and robustness have been recognized as key challenges for Surface-enhanced Raman scattering (SERS) analytical techniques. Quantifying the concentration of an analyte using a single characteristic peak from SERS has been a controversial topic because the Raman signal is susceptible to highly concentrated electromagnetic hotspots, inhomogeneity of SERS substrate, or non-standardization of measurement conditions. Ratiometric SERS strategies have been demonstrated as a promising solution to effectively balance and compensate for signal fluctuations caused by matrix heterogeneity. However, it is not easy to construct ratiometric SERS sensors with monitoring the ratio of two different signal intensities for target analysis. RESULTS: An attempt has been made to develop a novel ratiometric biosensor that can be applied to detect okadaic acid (OA). Aptamer-anchored magnetic particles were first combined with gold-tagged short complementary DNA (Au-cDNA) to create heterogeneous nanostructures. When the target was present, the Au-cDNA was dissociated from nanostructures, and 4-nitrothiophenol (4-NTP) was initiated to reduce to 4-aminothiophenol (4-ATP) in the presence of hydrogen sources. The SERS ratio change of 4-NTP and 4-ATP was finally detected by AuNPs-coated film. OA was successfully quantified, and the detection limit was as low as 2.4524 ng/mL. The constructed biosensor had good stability and reproducibility with a relative standard deviation of less than 4.47%. The proposed method used gold nanoparticles as an intermediate to achieve catalytic signal amplification and subsequently increased the sensitivity of the biosensor. SIGNIFICANCE AND NOVELTY: Catalytic reaction-based ratiometric SERS biosensors combine the multiple advantages of catalytic signal amplification and signal self-calibration and provide new insights into the development of stable, reproducible, and reliable SERS detection techniques. This ratiometric SERS technique offered a universal method that is anticipated to be applicable for the detection of other targets by substituting the aptamer.

Ag@Au core-shell nanoparticle-based surface-enhanced Raman scattering coupled with chemometrics for rapid determination of chloramphenicol residue in fish.

The alarming increase in drug-resistant bacteria in fish resulting from the misuse of antibiotics poses a significant threat to ecosystems and human health. Therefore, the development of a reliable approach for detecting antibiotic residues in fish is crucial. In this study, a rapid and simple method for detecting chloramphenicol (CAP) residue in tilapia was developed using surface-enhanced Raman scattering (SERS) combined with chemometric algorithms. Silver and gold core-shell nanoparticles (Ag@Au CSNPs) were used as SERS nanosensors to achieve strong signal amplification with an enhancement factor of 2.67 × 106. The results demonstrated that the variable combination population analysis-partial least square (VCPA-PLS) model combined with the standard normal variable transformation pretreatment method exhibited the best predictive performance with a detection limit of 1 × 10-5 µg/mL. Thus, an SERS technique was established based on Ag@Au CSNPs combined with VCPA-PLS to rapidly detect CAP in tilapia.

Isothermal Reciprocal Catalytic DNA Circuit for Sensitive Analysis of Kanamycin.

Inappropriate use of veterinary drugs can result in the presence of antibiotic residues in animal-derived foods, which is a threat to human health. A simple yet efficient antibiotic-sensing method is highly desirable. Programmable DNA amplification circuits have supplemented robust toolkits for food contaminants monitoring. However, they currently face limitations in terms of their intricate design and low signal gain. Herein, we have engineered a robust reciprocal catalytic DNA (RCD) circuit for highly efficient bioanalysis. The trigger initiates the cascade hybridization reaction (CHR) to yield plenty of repeated initiators for activating the rolling circle amplification (RCA) circuit. Then the RCA-generated numerous reconstituted triggers can reversely stimulate the CHR circuit. This results in a self-sufficient supply of numerous initiators and triggers for the successive cross-invasion of CHR and RCA amplifiers, thus leading to exponential signal amplification for the highly efficient detection of analytes. With its flexible programmability and modular features, the RCD amplifier can serve as a universal toolbox for the high-performance and accurate sensing of kanamycin in buffer and food samples including milk, honey, and fish, highlighting its enormous promise for low-abundance contaminant analysis in foodstuffs.

Metal-organic framework-based multienzyme cascade bioreactor for sensitive detection of methyl parathion.

In this study, a cascade nanobioreactor was developed for the highly sensitive detection of methyl parathion (MP) in food samples. The simultaneous encapsulation of acetylcholinesterase (AChE) and choline oxidase (CHO) in a zeolitic imidazole ester backbone (ZIF-8) effectively improved the stability and cascade catalytic efficiency of the enzymes. In addition, glutathione-stabilized gold nanoclusters (GSH-AuNCs) were encapsulated in ZIF-8 by ligand self-assembly, conferring excellent fluorescence properties. Acetylcholine (ATCh) is catalyzed by a cascade of AChE/CHO@ZIF-8 as well as Fe(II) to generate hydroxyl radicals (·OH) with strong oxidizing properties. The ·OH radicals then oxidize Au(0) in GSH-AuNCs@ZIF-8 to Au(I), resulting in fluorescence quenching. MP, as an inhibitor of AChE, hinders the cascade reaction and thus restores the fluorescence emission, enabling its quantitative detection. The limit of detection of the constructed nanobioreactor for MP was 0.23 µg/L. This MOF-based cascade nanobioreactor has great potential for the detection of trace hazards.

Sulfadiazine detection in aquatic products using upconversion nanosensor based on photo-induced electron transfer with imidazole ligands and copper ions.

Contamination of aquatic products with sulfonamide antibiotics poses a threat to consumer health and can lead to the emergence of drug-resistant bacteria. Common methods to detect such compounds are slow and require expensive instruments. We developed a sensitive sulfadiazine (SDZ) detection method based on the photoinduced electron transfer between UCNPs and Cu2+. The surface-modified upconversion nanoparticles bind to Cu2+ by electrostatic adsorption, causing fluorescence quenching. The quenched fluorescence was subsequently recovered by the addition of imidazole and SDZ to the detection system, which formed a complex with Cu2+. The sensor showed excellent linearity over a wide concentration range (0.05-1000 ng/mL), had a low limit of detection (0.04 ng/mL), was selective, and was not affected by common substances present in aquatic media. This indicates that the sensor has great potential for application in the detection of SDZ residues in aquatic products.

Upconversion fluorescence nanosensor based on enzymatic inhibited and copper-triggered o-phenylenediamine oxidation for the detection of dimethoate pesticides.

Pesticide residues in agricultural products pose a significant threat to human health. Herein, a sensitive fluorescence method employing upconversion nanoparticles was developed for detecting organophosphorus pesticides (OPs) based on the principle of enzyme inhibition and copper-triggered o-phenylenediamine (OPD) oxidation. Copper ions (Cu2+) oxidized the colorless OPD to a yellow 2,3-diaminophenazine (oxOPD). The yellow solution oxOPD quenched the fluorescence of upconversion nanoparticles due to the fluorescence resonance energy transfer. The high affinity of Cu2+ for thiocholine reduced the level of oxOPD, resulting in almost no fluorescence quenching. The addition of dimethoate led to the inhibition of acetylcholinesterase activity and thus prevented the formation of thiocholine. Subsequently, Cu2+ oxidized OPD to form oxOPD, which attenuated the fluorescence signal of the system. The detection system has a good linear range of 0.01 ng/mL to 50 ng/mL with a detection limit of 0.008 ng/mL, providing promising applications for rapid detection of dimethoate.

Rapid detection of eugenol in perch utilizing electrochemical method by transition metal substituted polyoxometalates.

Food safety concerns caused by the application of spice allergens to fish anaesthesia. In this paper, a chitosan-reduced graphene oxide/polyoxometalates/poly-l-lysine (CS-rGO/P2Mo17Cu/PLL) modified electrode was constructed by electrodeposition and successfully applied to the quantitative analysis of eugenol (EU). The detection limit was 0.4490 µM in the linear range of 2x10-6 M to 1.4x10-5 M. It was applied to the determination of EU residues in kidney, liver and meat tissues of perch with recoveries ranging from 85.43 to 93.60%. Besides, the electrodes demonstrate high stability (2.56% drop in current value after 70 days at room temperature), high reproducibility (RSD of 4.87% for 6 parallel electrodes) and extremely fast response time. This study provided a new material for the electrochemical detection of EU.

Screening-Capture-Integrated Electrochemiluminescent Aptasensor Based on Mesoporous Silica Nanochannels for the Ultrasensitive Detection of Deoxynivalenol in Wheat.

To prevent the contamination of cereals by mycotoxins, establishing a sensitive and rapid method for the detection of mycotoxins is essential. In this study, a screening-capture-integrated electrochemiluminescence (ECL) aptasensor based on mesoporous silica films (MSFs) was successfully prepared for the ultrasensitive and highly selective detection of deoxynivalenol (DON) in wheat. The narrow nanochannels of MSFs can realize size screening, thereby eliminating the influence of macromolecular substances and providing a pure environment for the signal probe (tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)32+)) to reach the indium tin oxide (ITO) conductive substrate, which significantly improves the anti-interference ability of the screening-capture-integrated ECL sensor. The aptamer (Apt) attached to the surface of the MSFs can specifically capture DON, and the resulting DON-Apt complex has a gated effect on the MSFs, triggering the inhibition of Ru(bpy)32+ in the electrolyte from reaching the ITO surface. Therefore, the ECL intensity of the sensor decreased with increasing DON concentration to achieve a quantitative detection of DON. Under optimized conditions, the linear range of the screening-capture-integrated ECL aptasensor was 0.001-200 µg/kg, and the detection limit was as low as 5.27 × 10-5 µg/kg (S/N = 3). In conclusion, this study developed a screening-capture-integrated ECL aptasensor that combines size screening and specific capture for the detection of DON in wheat, providing a new approach for the early detection of wheat mildew.

Facile synthesis of fluorescence-SERS dual-probe nanocomposites for ultrasensitive detection of sulfur-containing gases in water and beer samples.

A highly structured fluorescent-SERS dual-probe nanocomposites were synthesized for the determination of sulfur-containing gases in water and beer samples. Initially, Au@Ag NPs were prepared by growing the Ag shell on the Au core in situ, modified with surfactant and fabricated with Zn2+. Then, MOF-5-NH2 assembled Au@Ag NPs were obtained through coordination between Zn sites and 2-aminoterephthalic acid. The principle was based on redox reaction between H2S and Au@Ag NPs, and the fluorescence turn-on effects were due to the charge transfer between SO2 and amino groups. The SERS intensity was related to the concentration of H2S (5 âˆ¼ 60 nM), and an ultra-low detection limit of 2.26 nM was achieved. Importantly, the fluorescence performance was applied for SO2 analysis and exhibited good linear response. Moreover, the platform for H2S and SO2 in real samples revealed satisfactory results (95.6 âˆ¼ 101.6% and 99.0 âˆ¼ 104.4%). Therefore, the proposed system offered a precise detection of H2S/SO2 in food/environmental settings.

Label-free detection of trace level zearalenone in corn oil by surface-enhanced Raman spectroscopy (SERS) coupled with deep learning models.

Surface-enhanced Raman spectroscopy (SERS) and deep learning models were adopted for detecting zearalenone (ZEN) in corn oil. First, gold nanorods were synthesized as a SERS substrate. Second, the collected SERS spectra were augmented to improve the generalization ability of regression models. Third, five regression models, including partial least squares regression (PLSR), random forest regression (RFR), Gaussian progress regression (GPR), one-dimensional convolutional neural networks (1D CNN), and two-dimensional convolutional neural networks (2D CNN), were developed. The results showed that 1D CNN and 2D CNN models possessed the best prediction performance, i.e., determination of prediction set (RP2) = 0.9863 and 0.9872, root mean squared error of prediction set (RMSEP) = 0.2267 and 0.2341, ratio of performance to deviation (RPD) = 6.548 and 6.827, limit of detection (LOD) = 6.81 × 10-4 and 7.24 × 10-4 µg/mL. Therefore, the proposed method offers an ultrasensitive and effective strategy for detecting ZEN in corn oil.

On-line monitoring of total sugar during kombucha fermentation process by near-infrared spectroscopy: Comparison of linear and non-linear multiple calibration methods.

Kombucha is widely recognized for its health benefits, and it facilitates high-quality transformation and utilization of tea during the fermentation process. Implementing on-line monitoring for the kombucha production process is crucial to promote the valuable utilization of low-quality tea residue. Near-infrared (NIR) spectroscopy, together with partial least squares (PLS), backpropagation neural network (BPANN), and their combination (PLS-BPANN), were utilized in this study to monitor the total sugar of kombucha. In all, 16 mathematical models were constructed and assessed. The results demonstrate that the PLS-BPANN model is superior to all others, with a determination coefficient (R2p) of 0.9437 and a root mean square error of prediction (RMSEP) of 0.8600 g/L and a good verification effect. The results suggest that NIR coupled with PLS-BPANN can be used as a non-destructive and on-line technique to monitor total sugar changes.

Boric acid group-functional Tb-MOF as a fluorescent and captured probe for the highly sensitive and selective determination of propyl gallate in edible oils.

This study reports the development of a Tb-metal-organic framework (Tb-MOF)-based fluorescent platform for the detection of propyl gallate (PG). The Tb-MOF using 5-boronoisophthalic acid (5-bop) as the ligand exhibited multiple emissions at 490, 543, 585, and 622 nm under an excitation wavelength of 256 nm. The fluorescence of Tb-MOF was selectively and significantly weakened in the presence of PG due to the special nucleophilic reaction between the boric acid of Tb-MOF and o-diphenol hydroxyl of PG, and the combined effect of static quenching and internal filtering. Furthermore, this sensor enabled the determination of PG within seconds in a wide linear range of 1-150 µg/mL, and with a low detection limit of 0.098 µg/mL, and high specificity against other phenolic antioxidants. This work provided a new route for the sensitive and selective determination of PG in soybean oil, thus was perspective to monitor and reduce the risk of PG overuse.

Construction of a self-sufficient DNA circuit for amplified detection of kanamycin.

Improper use of kanamycin can lead to trace kanamycin residues in animal-derived foods, which can pose a potential threat to public health. Isothermal enzyme-free DNA circuits have provided a versatile toolbox for detecting kanamycin residues in complicated food samples, yet they are always limited by low amplification efficiency and intricate design. Herein, we present a simple-yet-robust nonenzymatic self-driven hybridization chain reaction (SHCR) amplifier for kanamycin determination with 5800-fold sensitivity over that of the conventional HCR circuit. The analyte kanamycin-activated SHCR circuitry can generate numerous new initiators to promote the reaction and improve the amplification efficiency, thus achieving an exponential signal gain. With precise target recognition and multilayer amplification capability, our self-sustainable SHCR aptasensor facilitated the highly sensitive and reliable analysis of kanamycin in buffer, milk, and honey samples, thus holding great potential for the amplified detection of trace contaminants in liquid food matrices.

G-Quadruplex/Hemin-Mediated Polarity-Switchable and Photocurrent-Amplified System for Escherichia coli O157:H7 Detection.

The contamination of food by pathogens is a serious problem in global food safety, and current methods of detection are costly, time-consuming, and cumbersome. Therefore, it is necessary to develop rapid, portable, and sensitive assays for foodborne pathogens. In addition, assays for foodborne pathogens must be resistant to interference resulting from the complex food matrix to prevent false positives and negatives. In this study, hemin and reduced graphene oxide-MoS2 sheets (GMS) were used to design a near-infrared (NIR)-responsive photoelectrochemical (PEC) aptasensor with target-induced photocurrent polarity switching based on a hairpin aptamer (Hp) with a G-quadruplex motif. A ready-to-use analytical device was developed by immobilizing GMS on the surface of a commercial screen-printed electrode, followed by the attachment of the aptamer. In the presence of Escherichia coli O157:H7, the binding sites of Hp with the G-quadruplex motif were opened and exposed to hemin, leading to the formation of a G-quadruplex/hemin DNAzyme. Crucially, after binding to hemin, the charge transfer pathway of GMS changes, resulting in a switch of the photocurrent polarity. Further, G-quadruplex/hemin DNAzyme enhanced the cathodic photocurrent, and the proposed sensor exhibited a wide linear range ((25.0-1.0) × 107 CFU/mL), a low limit of detection (2.0 CFU/mL), and good anti-interference performance. These findings expand the applications of NIR-responsive PEC materials and provide versatile PEC methods for detecting biological analytes, especially for food safety testing.

Electrochemical sensing of nitrofurazone on Ru(bpy)32+ functionalized polyoxometalate combined with graphene modified electrode.

Nitrofurazone is forbidden to be used in aquaculture, but it is often used illegally because of its good bactericidal effect, and its content in animals is extremely low and difficult to detect directly. Hence, a functionalized polyoxometalate combined with graphene modified electrodes through layer-by-layer assembly has achieved a sensitive detection of nitrofurazone in a pH = 6 Na2HPO4-citrate buffer solution and its detection limit as low as 0.08952 µM. Nitrofurazone has accelerated its electron transfer through [Ru-PMo12/PDDA-GO]3 modified electrode, thus realizing its direct detection at low levels through actual samples. This study provides a new perspective for the direct detection of nitrofurazone by electrochemical methods, which is of great significance for the supervision of nitrofurazone and the improvement of the quality and safety of aquatic products.