A dual-recognition strategy is reported to construct a one-step washing and highly efficient signal-transduction tag system for high-sensitivity colorimetric detection of Staphylococcus aureus (S. aureus). The porous (gold core)@(platinum shell) nanozymes (Au@PtNEs) as the signal labels show highly efficient peroxidase mimetic activity and are robust. For the sake of simplicity the detection involved the use of a vancomycin-immobilized magnetic bead (MB) and aptamer-functionalized Au@PtNEs for dual-recognition detection in the presence of S. aureus. In addition, we designed a magnetic plate to fit the 96-well microplate to ensure consistent magnetic properties of each well, which can quickly remove unreacted Au@PtNEs and sample matrix while avoiding tedious washing steps. Subsequently, Au@PtNEs catalyze hydrogen peroxide (H2O2) to oxidize 3,3\',5,5\'-tetramethylbenzidine (TMB) generating a color signal. Finally, the developed Au@PtNEs-based dual-recognition washing-free colorimetric assay displayed a response in the range of S. aureus of 5 × 101-5 × 105 CFU/mL, and the detection limit was 40 CFU/mL within 1.5 h. In addition, S. aureus-fortified samples were analyzed to further evaluate the performance of the proposed method, which yielded average recoveries ranging from 93.66 to 112.44% and coefficients of variation (CVs) within the range 2.72-9.01%. These results furnish a novel horizon for the exploitation of a different mode of recognition and inexpensive enzyme-free assay platforms as an alternative to traditional enzyme-based immunoassays for the detection of other Gram-positive pathogenic bacteria.

UNASSIGNED: Ultrasensitive bacterial detection methods are crucial to ensuring accurate diagnosis and effective clinical monitoring, given the significant threat bacterial infections pose to human health. The aim of this study is to develop a biosensor with capabilities for broad-spectrum bacterial detection, rapid processing, and cost-effectiveness.
UNASSIGNED: A magnetically-assisted SERS biosensor was designed, employing wheat germ agglutinin (WGA) for broad-spectrum recognition and antibodies for specific capture. Gold nanostars (AuNSs) were sequentially modified with the Raman reporter molecules and WGA, creating a versatile SERS tag with high affinity for a diverse range of bacteria. Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) antibody-modified Fe3O4 magnetic gold nanoparticles (MGNPs) served as the capture probes. Target bacteria were captured by MGNPs and combined with SERS tags, forming a \"sandwich\" composite structure for bacterial detection.
UNASSIGNED: AuNSs, with a core size of 65 nm, exhibited excellent storage stability (RSD=5.6%) and demonstrated superior SERS enhancement compared to colloidal gold nanoparticles. Efficient binding of S. aureus and P. aeruginosa to MGNPs resulted in capture efficiencies of 89.13% and 85.31%, respectively. Under optimized conditions, the developed assay achieved a limit of detection (LOD) of 7 CFU/mL for S. aureus and 5 CFU/mL for P. aeruginosa. The bacterial concentration (10-106 CFU/mL) showed a strong linear correlation with the SERS intensity at 1331 cm-1. Additionally, high recoveries (84.8% - 118.0%) and low RSD (6.21% - 11.42%) were observed in spiked human urine samples.
UNASSIGNED: This study introduces a simple and innovative magnetically-assisted SERS biosensor for the sensitive and quantitative detection of S. aureus or P. aeruginosa, utilizing WGA and antibodies. The developed biosensor enhances the capabilities of the \"sandwich\" type SERS biosensor, offering a novel and effective platform for accurate and timely clinical diagnosis of bacterial infections.

In this study, we reported the eco-responsible synthesis of iron-doped carbon quantum dots (Fe-CQDs) from waste coffee grounds through a simple hydrothermal method. The Fe-CQDs exhibited high peroxidase-like activity, which could convert 3,3\',5,5\'-tetramethylbenzidine (TMB) into blue ox-TMB in the presence of H2O2. After adding ascorbic acid (AA) to above system, the blue solution faded. Based on this phenomenon, a colorimetric method for visual monitoring of H2O2 and AA was developed. Meanwhile, the fluorescence of Fe-CQDs can be quenched by the formed ox-TMB via inner filter effect (IFE), followed by the recovery upon the addition of AA. Therefore, Fe-CQDs can be acted as a fluorescent probe to detect H2O2 and AA through the \"on-off-on\" mode. Furthermore, the dual-recognition methods based on Fe-CQDs were used to measure AA content in beverage samples. Thus, this work would shed much light on converting waste into biomass CQDs and their potential applications in biomolecular detection.

We described a two-step förster resonance energy transfer (FRET) system for ratiometric Staphylococcus aureus (S. aureus) detection based on a dual-recognition proximity binding-induced toehold strand displacement reactions (TSDR). Ru(bpy)32+ and platinum nanoparticles (Pt NPs) labeled DNA (Ru-S3 and Pt NPs-S4) hybridized to enable the occurrence of the primary FRET using Ru(bpy)32+ as the energy donor and Pt NPs as the energy acceptor. TSDR happened by integrating vancomycin hydrochloride labeled S1 (Van-S1) and gold nanoclusters labeled S2-aptamer (Au NCs-S2-aptamer) with S. aureus. The single DNA segments of Van-S1 bond to the terminal toehold of Ru-S3, displacing Pt-S4, inducing the secondary FRET using Au NCs as the energy donor and Ru(bpy)32+ as the energy acceptor. This two-step FRET system efficiently improved the reaction efficiency of S. aureus with a detection limit of 1.0 CFU/mL. Furthermore, satisfactory results obtained while detecting S. aureus in food samples, indicating a great potential for food analysis.

Rapid, sensitive and simultaneous detection of multiple bacteria in foodborne is still a major challenge in public health field. Here, a fluorescence immunoassay that can achieve high-throughput detection of three Gram-positive foodborne pathogenic bacteria simultaneously was proposed. Vancomycin and bovine serum albumin conjugate (Van-BSA) was immobilized on a polycarbonate chip to capture three Gram-positive foodborne pathogenic bacteria, Staphylococcus aureus (S. aureus), Bacillus cereus (B. cereus) and Listeria monocytogenes (L. monocytogenes). CdSe/ZnS quantum dot modified antibodies (Ab-QD) were prepared by carbodiimide coupling chemistry. Due to the affinity reaction between antibodies and proteins on the bacterial surface, the simultaneous detection of multiple Gram-positive bacteria was achieved by monitoring the fluorescence signal of quantum dot by a portable microfluidic chip analyzer. Under optimal conditions, low detection limits was 18 CFU/well, 3 CFU/well and 36 CFU/well for S. aureus, B. cereus and L. monocytogenes, respectively. With satisfactory accuracy and precision, the proposed fluorescence immunoassay holds good prospects to detect pathogens in real food samples.

Circulating tumor cells (CTCs) are promising liquid biopsy biomarkers for early cancer detection and anti-cancer therapy evaluation. The ultra-low abundance of CTCs in blood samples requires highly sensitive and accurate detection ways. In this study, we propose the design of a dual-recognition electrochemical biosensor to improve both the specificity and signal response. PdPtCuRu mesoporous nanospheres (PdPtCuRu MNSs) with excellent three dimensions (3D) nanopore structures were synthesized by one-pot method and connected to mucin 1 (MUC1) aptamer to serve as signal amplification probe. Besides, superconductive carbon black, Ketjen Black (KB), and gold nanoparticles (AuNPs) modified organometallic frame (CeMOF-Au) were combined to work as signal transducer. The characteristic branching structure of KB provides abundant contact points to load CeMOF-Au to heighten the interface electron transfer rate. In addition, AuNPs were reduced on the surface of CeMOF, which could effectively bind the capture antibody and further enhance the conductivity. Under the optimized condition, the limit of detection (LOD) of the as-constructed biosensor was less than 10 cells mL-1 for model A549 cells, and showed good specificity and accuracy in spiked serum samples. We envision the as-proposed electrochemical biosensor would alternate as a useful tool for the clinical detection of CTCs for cancer diagnosis.

the accurate, reliable and specific analysis of foodborne pathogenic bacteria is vital for human health and safety. Staphylococcus aureus (S. aureus), as a common bacterium, is regularly found in food, water, and other biological samples. Herein, a signal-off electrochemical DNA sensor (E-DNA sensor) was designed for the sensitive detection ofS. aureusamplified withthecombination of a dna walker and pb2+-specific dnazyme. In this work, vancomycin functionalized gold nanoclusters (Van@Au NCs) and an aptamer strand as identification units were modified at the termini of two proximity probes. upon the addition of targetS. aureus, a dual-recognition binding-induced dna walker was driven by the formation of pba dual-recognition binding-induced dna walker was driven by the formation of pba dual-recognition binding-induced dna walker was driven by the formation of pba dual-recognition binding-induced dna walker was driven by the formation of pb2+-dependent dnazyme, achieving the conversion of oneS. aureus to many intermediate dna (t) strands. then, the released t strands hybridized with methylene blue-tagged hairpin dna (h-mb) on the electrode. consequently, the conformational alteration of t strands reduced the electron transfer efficiency of mb to the electrodeinterface (signal-off). therefore, sensitive analysis of S. aureus was readily acquired within a range of 10-107 CFU/mL and a low detection limit at 1 CFU/mL. Undoubtedly, dual recognition by aptamer and vancomycin in an integrated scheme brought about a good recognition performance of S. aureus in complex samples, as well as an efficient annihilation of harmful pathogenic bacteria during the experiment.

Development of a simple and efficient dual-mode analytical technique with the built-in cross reference correction feature is benefit to achieve the highly accurate detection of the target pollutants and avoid the false-positive outputs in environmental media. Here, we synthesized a Fe-doped polydopamine (Fe@PDA)-based nanozyme with prominent peroxide-mimetic enzyme activity and high fluorescence emission ability. On this basis, we designed a dual-recognition strategy-driven fluorescence-colorimetric dual-mode detection platform, consisting of Listeria monocytogenes (L. monocytogenes) recognition aptamer-modified Fe@PDA (apt/Fe@PDA) and vancomycin-functionalized Fe3O4 (van/Fe3O4), for L. monocytogenes. Owing to van/Fe3O4-powered magnetic separation, there was a L. monocytogenes concentration-dependent fluorescence enhancement of apt/Fe@PDA for performing fluorescence assay in the precipitate. In this case, the prominent peroxide-mimetic enzyme activity of the residual apt/Fe@PDA in the precipitation could catalyze H2O2 to further oxidate colorless 3,3\',5,5\'-tetramethylbenzidine (TMB) into blue oxTMB, which displayed a L. monocytogenes concentration-dependent absorbance enhancement for carrying out colorimetric assay as well. As a result, a fluorescence-colorimetric dual-mode analytical platform was proposed to successfully detect the residual L. monocytogenes in real environmental media with acceptable results. This work showed the great prospects by integrating dual-recognition strategy into fluorescence nanozyme to develop efficient and reliable dual-mode analytical platforms for safeguarding environmental health.

A novel surface-enhanced Raman scattering (SERS)-based analytical technique was proposed to simultaneously detect two highly pathogenic bacteria, namely, Staphylococcus aureus (S. aureus) and Listeria monocytogenes (L. mono) by using a dual-recognition pattern with wheat germ agglutinin (WGA) and nucleic acid aptamers. WGA was modified onto Fe3O4@Au magnetic nanoparticles (MNPs) for the efficient capture of S. aureus and L. mono in complex samples (orange juice, extracts of lettuce, and human urine) within 15 min. The streptavidin (SA)/aptamers co-functionalized SERS tags were fabricated by covalent attaching two different Raman reporters and SA molecules onto 45 nm Au NPs and then conjugated with two biotin-aptamers that specifically bind to their target bacteria with high affinity and stability. The combined use of high-sensitive SERS tags, WGA-mediated magnetic enrichment, and SA-mediated aptamer conjugation remarkably improved the assay sensitivity. Under optimized conditions, the developed SERS biosensor can simultaneously detect the two target bacteria with high detection sensitivity (<6 cells/mL), favorable linear relation (10-107 cells/mL), and high accuracy (recovery rate <7.03%). Therefore, the proposed SERS platform is rapid, sensitive, easy to use, and thus show potential as a tool for the timely identification of pathogenic bacteria in real samples.

In this study, surface imprinting, magnetic separation, and fluorescent detection were integrated to develop a dual-recognition sensor (MF-MIPs), which was used for highly selective and sensitive detection of 4-nitrophenol (4-NP) in food samples. Silane-functionalized carbon dots (Si-CDs) participated in the imprinting process and were uniformly distributed into the MIPs layers. MF-MIPs sensor exhibited a high fluorescence response and selectivity based on the dual-recognition mechanism of imprinting recognition and fluorescence identification. The relative fluorescence intensity of MF-MIPs sensor presented a good linear relationship in the range of 0.08-10 μmol·L-1 with a low limit of detection (23.45 nmol·L1) for 4NP. MF-MIPs sensor showed high anti-interference, as well as excellent stability and reusability. The 4-NP recovery from spiked food samples ranged from 93.20 to 102.15%, and the relative standard deviation was lower than 5.0%. Therefore, MF-MIPs sensor may be a promising method for 4-NP detection in food samples.