Flower-like gold nanoparticles labeled and silver deposition rapid vertical flow technology for highly sensitive detection of Brucella antibodies.

To prevent the transmission of brucellosis, rapid vertical flow technology (RVFT) was developed to detect brucellosis antibodies. To improve the sensitivity of the technique, lipopolysaccharides (LPS) were purified and used to detect brucellosis antibodies. To improve the sensitivity of serum antibody detection, a single multifunctional buffer was established in whole blood and other biological samples, and the advantages of the lateral flow immunoassay were retained. Flower-like gold nanoparticles were applied to RVFT for the first time. In this study, silver ions were catalyzed by flower-like gold nanoparticles into metal silver deposited on the surface of gold nanoparticles for the first time, which not only increased the particle size of gold nanoparticles, but also showed a more distinguishable black color on the test zone, further improving the sensitivity of RVFT. Standard Brucella-positive serum (containing Brucella antibody at 4000 IU mL-1) could be detected in this system even for a dilution factor of 2 × 10-3. The detection limit was 2 IU mL-1. RVFT can effectively avoid the false negative phenomenon in lateral flow immunoassay. RVFT is simple to operate, with a short reaction time, 2-3 minutes visible to the naked eye, without any equipment. Because it is very important to control the brucellosis epidemic, this approach has great application prospects in basic medical units and for veterinarians.

Gold Nanoparticles Prepared with Cyclodextrin Applied to Rapid Vertical Flow Technology for the Detection of Brucellosis.

Currently, brucellosis seriously threatens the health of humans and animals and hinders the development of animal husbandry. However, the diagnostic methods for brucellosis have some disadvantages, such as low sensitivity, long detection time, professional operation, and high cost. This study aims to establish a convenient, fast, effective, and inexpensive detection method for brucellosis. Gold nanoparticles with ß-cyclodextrin as a reducing agent were prepared and optimized, applied to rapid vertical flow technology (RVFT), and used to establish a kit for the detection of brucellosis. In this study, gold nanoparticles prepared from ß-cyclodextrin were applied to RVFT for the first time, and on this basis, silver staining amplification technology was introduced, which further improved the sensitivity and reduced the detection limit of this method. Standard Brucella-Positive Serum (containing Brucella antibody at 4000 IU/mL) could be detected in this system even for a dilution factor of 1 × 10-3. The detection limit was 4 IU/mL. RVFT is simple to operate, has a short reaction time, and is 5-6 min visible to the naked eye, without any equipment.

Synthesis of Eco-Friendly Silver Nanoparticles Using Glycyrrhizin and Evaluation of Their Antibacterial Ability.

In the present study, the biosynthesis of silver nanoparticles (AgNPs) and their antibacterial activity against gram-positive and gram-negative bacteria were investigated. Glycyrrhizin (GL) was used as a reducing agent and stabilizer to rapidly prepare the AgNPs. The distinctive absorption peak at 419 nm confirmed the formation of GL-reduced AgNPs. The TEM and particle size analysis shows that the prepared GL-reduced AgNPs were mostly circular with good dispersion and a relatively uniform particle size of 35 nm on average. Fourier transform infrared spectroscopy analysis was performed to identify the possible biomolecules in the capping and active stabilization of the GL-reduced AgNPs. The antibacterial activity of the GL-reduced AgNPs was analyzed with the Oxford cup diffusion method and filter paper diffusion method. The experimental results show that these properties endowed the GL-reduced AgNPs with high antibacterial activity against Escherichia coli and Staphylococcus aureus and lay a foundation for the use of colloidal silver in antibacterial applications. The GL-reduced AgNPs also had stronger antibacterial activity than sodium citrate-reduced AgNPs, which indicates the advantages of GL-reduced AgNPs compared with sodium citrate-reduced AgNPs in inducing bacteriostasis. The cytotoxicity of GL-reduced AgNPs on human kidney epithelial 293A (HEK293) cells was evaluated via the MTT assay. The results show that GL-reduced AgNPs had lower toxicity to HEK293 cells than sodium citrate-AgNPs, which indicates that the as-prepared GL-reduced AgNPs are environmentally friendly.

Detection and Difference Analysis of the Enzyme Activity of Colloidal Gold Nanoparticles With Negatively Charged Surfaces Prepared by Different Reducing Agents.

Nanozymes are particles with diameters in the range of 1-100 nm, which has been widely studied due to their biological enzyme-like properties and stability that natural enzymes do not have. In this study, several reducing agents with different structures (catechol (Cc), hydroquinone (Hq), resorcinol (Rs), vitamin C (Vc), pyrogallic acid (Ga), sodium citrate (Sc), sodium malate (Sm), and sodium tartrate (St)) were used to prepare colloidal gold with a negative charge and similar particle size by controlling the temperature and pH. The affinity analysis of the substrate H2O2 and TMB showed that the order of activities of colloidal gold Nanozymes prepared by different reducing agents was Cc, Hq, Rs, Vc, Ga, Sc, Sm, St. It was also found that the enzyme activity of colloidal gold reduced by benzene rings is higher than that of the colloidal gold enzyme reduced by linear chains. Finally, we discussed the activity of the colloidal gold peroxidase based on the number and position of isomers and functional groups; and demonstrated that the nanozymes activity is affected by the surface activity of colloidal gold, the elimination of hydroxyl radicals and the TMB binding efficiency.

Green synthesis and characterization of gold nanoparticles and their application for the rapid detection of glycyrrhizin with immunochromatographic strips.

In this study, a facile and environmentally friendly synthesis process was proposed without regular chemical additives. We successfully synthesized spherical gold nanoparticles (AuNPs) coated with glycyrrhizin (GL) by using GL as both a reductant and a stabilizer to reduce chloroauric acid. The obtained NPs were approximately 35 nm in size. The formation of these GL-AuNPs was verified by the presence of a surface plasmon resonance band at 526 nm. We also experimentally determined that in terms of chemical structure, GL can be used as a reducing agent to obtain colloidal gold. The d-glucuronic acid structure, rather than glycyrrhetinic acid (GA), plays an important reducing role in colloidal gold production. From this, we hypothesized that other compounds with sugar structures in Glycyrrhiza may also have the ability to reduce chloroauric acid. To mitigate the high cost and low efficiency of current GL detection methods, we applied AuNPs to the immunochromatographic system. Then, a colloidal gold immunochromatographic test strip based on the indirect competition method was developed for the rapid detection of GL, and the detection limit of this strip was 25 ng mL-1. The cross-test showed that the strip has high specificity. The test results are consistent with the data obtained by high-performance liquid chromatography (HPLC) with a coincidence rate of up to 100%. The rapid test strip is simple, fast, highly efficient and inexpensive, making it suitable for large-scale, rapid glycyrrhizin content determination.