ABSTRACT
Klebsiella pneumoniae, a member of the family Enterobacteriaceae, is a rod-shaped, Gram-negative bacterium, mainly found in the hospital environment and medical tools. It is the leading cause of nosocomial infection, characterized by bloodstream infection, wound site infection, urinary tract infection, and sepsis, mostly in older adults, newborn infants, and immunocompromised patients. This present study demonstrated a novel diagnostic method for K. pneumoniae detection based on the gold nanozyme activity for the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. The nanozyme activity of AuNPs with staining enhancement was statistically three times higher than that of the bare AuNPs in solid absorption at 650 nm. Nano-ELISA with staining enhancement could detect as low as 102 CFUs/mL of K. pneumoniae concentration, as the cutoff value was determined to be 0.158, which boosted the sensitivity of the immunoreactions by up to 100-fold. The detection limit of our assays was 26.023 CFUs/mL, and the limit of quantification was 78.857 CFUs/mL. There was no cross-reaction against other bacteria, which proved the immunoassays' remarkable specificity for recognizing K. pneumoniae. Taken together, we successfully developed and optimized the highly sensitive and decently specific nano-ELISA strategy that might be applicable for detecting various other bacterial pathogens.
ABSTRACT
The benefits of molecularly imprinted polymer (MIP) technology in creating artificial receptors to replace natural counterparts have piqued the interest of numerous researchers in recent years. We propose a biomimetic enrofloxacin-MIP for enrofloxacin (ENRO) antigen detection using gold nanoparticles (AuNPs) and MIP methodologies in this study. A self-assembled monomer layer of aminothiophenol was used to immobilize a pre-formed complex of the anti-enrofloxacin monoclonal antibody and enrofloxacin antigen onto the surface of an AuNP coated screen-printed carbon ink electrode (SPCE). The poly-(aminothiophenol) layer thickness was adjusted to entrap and restrict enrofloxacin antigens near the surface. The imprinting and removal of the enrofloxacin antigen in the MIP film were strongly validated by the Raman spectra. The final mAb-MIP sensor had better sensitivity (302 Ω mL ng-1) and a better detection limit (0.05 ng mL-1) than self-assembled monolayer (SAM)-based immunosensors, which had 102 Ω mL ng-1 and 0.1 ng mL-1, respectively.
Subject(s)
Biosensing Techniques , Metal Nanoparticles , Molecular Imprinting , Biomimetics , Biosensing Techniques/methods , Electrochemical Techniques/methods , Enrofloxacin , Gold , Immunoassay , Molecular Imprinting/methods , Molecularly Imprinted Polymers , PolymersABSTRACT
An innovative novel interface has been designed and developed to be used as a potential active layer in chemically sensitive field-effect transistor (Chem-FET) sensor devices for the specific sensing of Cs+. In this study, the synthesis of a specific Cs+ probe based on calix[4]arene benzocrown ether, its photophysical properties, and its grafting onto a single lipid monolayer (SLM) recently used as an efficient ultrathin organic dielectric in Chem-FETs are reported simultaneously. On the basis of both optical and NMR titration experiments, the probe has shown high selectivity and specificity for Cs+ compared to interfering cations, even if an admixture is used. Additionally, Attenuated Total Reflectance Fourier Transform Infra Red (ATR-FTIR) spectroscopy was successfully used to characterize and prove the efficient grafting of the probe onto a SLM and the formation of the innovative novel sensing layer.
