Selective and sensitive detection of lysozyme based on plasmon resonance light-scattering of hydrolyzed peptidoglycan stabilized-gold nanoparticles.

The simple, economic, rapid, and sensitive detection of lysozyme has an important significance for disease diagnosis since it is a potential biomarker. In this work, a new detection strategy for lysozyme was developed based on the change of the plasmon resonance light scattering (PRLS) signal of peptidoglycan stabilized gold nanoparticles (PGN-AuNPs). Peptidoglycan (PGN) was employed as a stabilizer to prepare PGN-AuNPs which have the properties of a uniform particle size, good stability, and a specific biological function. Due to the specific cleavage of lysozyme to PGN, a very simple specific and sensitive detection method for lysozyme was developed based on the PRLS signal of PGN-AuNPs after mixing with lysozyme for 1.5 h. The enhanced PRLS signals (ΔIPRLS, at 560 nm) increased linearly with increasing lysozyme in the range 5 nM to 1600 nM with the detection limit down to 2.32 nM (ΔIPRLS = 41.6397 + 0.5332c, R = 0.9961). When the PGN-AuNP based method was applied to assay lysozyme in authentic human serum samples, the recovery efficiency was 106.76-119.32% with the relative standard deviations in the range of 0.14-3.11%, showing good feasibility. The PGN-AuNP based method for lysozyme assay developed here is simple, rapid, selective, and sensitive, which is expected to provide a feasible new method for the diagnosis or prognosis of lysozyme-related diseases in a clinical setting.

Dual-Recognition Förster Resonance Energy Transfer Based Platform for One-Step Sensitive Detection of Pathogenic Bacteria Using Fluorescent Vancomycin-Gold Nanoclusters and Aptamer-Gold Nanoparticles.

The effective monitoring, identification, and quantification of pathogenic bacteria is essential for addressing serious public health issues. In this study, we present a universal and facile one-step strategy for sensitive and selective detection of pathogenic bacteria using a dual-molecular affinity-based Förster (fluorescence) resonance energy transfer (FRET) platform based on the recognition of bacterial cell walls by antibiotic and aptamer molecules, respectively. As a proof of concept, Vancomycin (Van) and a nucleic acid aptamer were employed in a model dual-recognition scheme for detecting Staphylococcus aureus (Staph. aureus). Within 30 min, by using Van-functionalized gold nanoclusters and aptamer-modified gold nanoparticles as the energy donor and acceptor, respectively, the FRET signal shows a linear variation with the concentration of Staph. aureus in the range from 20 to 108 cfu/mL with a detection limit of 10 cfu/mL. Other nontarget bacteria showed negative results, demonstrating the good specificity of the approach. When employed to assay Staph. aureus in real samples, the dual-recognition FRET strategy showed recoveries from 99.00% to the 109.75% with relative standard derivations (RSDs) less than 4%. This establishes a universal detection platform for sensitive, specific, and simple pathogenic bacteria detection, which could have great impact in the fields of food/public safety monitoring and infectious disease diagnosis.

Antibiotics mediated facile one-pot synthesis of gold nanoclusters as fluorescent sensor for ferric ions.

In this paper, the cheap, easily obtained small antibiotic molecule of vancomycin was employed as reducer/stabilizer for facile one-pot synthesis of water exhibited a bluish fluorescence emission at 410nm within a short synthesis time about 50min. Based on the strong fluorescence quenching due to electron transfer mechanism by the introduction of ferric ions(Fe3+), the Van-AuNCs were interestingly designed for sensitive and selective detecting Fe3+ with a limit of 1.4µmol L-1 in the linear range of 2-100µmol L-1 within 20min. The Van-AuNCs based method was successfully applied to determine Fe3+ in tap water, lake water, river water and sea water samples with the quantitative spike recoveries from 97.50-111.14% with low relative standard deviations ranging from 0.49-1.87%, indicating the potential application of this Van-AuNCs based fluorescent sensor for environmental sample analysis.

Dual Recognition Strategy for Specific and Sensitive Detection of Bacteria Using Aptamer-Coated Magnetic Beads and Antibiotic-Capped Gold Nanoclusters.

Food poisoning and infectious diseases caused by pathogenic bacteria such as Staphylococcus aureus (SA) are serious public health concerns. A method of specific, sensitive, and rapid detection of such bacteria is essential and important. This study presents a strategy that combines aptamer and antibiotic-based dual recognition units with magnetic enrichment and fluorescent detection to achieve specific and sensitive quantification of SA in authentic specimens and in the presence of much higher concentrations of other bacteria. Aptamer-coated magnetic beads (Apt-MB) were employed for specific capture of SA. Vancomycin-stabilized fluorescent gold nanoclusters (AuNCs@Van) were prepared by a simple one-step process and used for sensitive quantification of SA in the range of 32-10(8) cfu/mL with the detection limit of 16 cfu/mL via a fluorescence intensity measurement. And using this strategy, about 70 cfu/mL of SA in complex samples (containing 3 × 10(8) cfu/mL of other different contaminated bacteria) could be successfully detected. In comparison to prior studies, the developed strategy here not only simplifies the preparation procedure of the fluorescent probes (AuNCs@Van) to a great extent but also could sensitively quantify SA in the presence of much higher concentrations of other bacteria directly with good accuracy. Moreover, the aptamer and antibiotic used in this strategy are much less expensive and widely available compared to common-used antibodies, making it cost-effective. This general aptamer- and antibiotic-based dual recognition strategy, combined with magnetic enrichment and fluorescent detection of trace bacteria, shows great potential application in monitoring bacterial food contamination and infectious diseases.

Multi-color quantum dot-based fluorescence immunoassay array for simultaneous visual detection of multiple antibiotic residues in milk.

Antibiotic residues, which are among the most common contaminants in animal-based food products such as milk, have become a significant public health concern. Here, we combine a multicolor quantum dot (QD)-based immunofluorescence assay and an array analysis method to achieve simultaneous, sensitive and visual detection of streptomycin (SM), tetracycline (TC), and penicillin G (PC-G) in milk. Antibodies (Abs) for SM, TC and PC-G were conjugated to QDs with different emission wavelengths (QD 520 nm, QD 565 nm and QD 610 nm) to serve as detection probes (QD-Ab). Then a direct competitive fluorescent immunoassay was performed in antigen-coated microtiter plate wells for simultaneous qualitative and quantitative detection of SM, TC, and PC-G residues, based on fluorescence of the QD-Ab probes. The linear ranges for SM, TC and PC-G were 0.01-25 ng/mL, 0.01-25 ng/mL and 0.01-10 ng/mL, respectively, with detection limit of 5 pg/mL for each of them. Based on fluorescence of the QD-Ab probes, residues of the three antibiotics were determined visually and simultaneously. Compared with a commercial enzyme-linked immunosorbent assay kit, our method could achieve simultaneous analysis of multiple target antibiotics in multiple samples in a single run (high-throughput analysis) and improved accuracy and sensitivity for analysis of residues of the three antibiotics in authentic milk samples. This new analytical tool can play an important role in ameliorating the negative impact of the residual antibiotics on human health and the ecosystem.