ABSTRACT
A label-free DNA hybridization electrochemical sensor for the detection of Klebsiella pneumoniae was developed, which could be helpful in the diagnosis of bacterial infections. Indole-5-carboxylic acid (ICA) and graphene oxide (GO) were electrodeposited on a glassy carbon electrode, and the resulting reduced GO (rGO)-ICA hybrid film served as a platform for immobilizing oligonucleotides on a single-stranded DNA (ssDNA) sequence. The conditions were optimized, with excellent electrochemical performance. A significant change was observed after hybridization of ssDNA with the target probe under optimum conditions. Hybridization with complementary, noncomplementary, one-base mismatched, and three-base mismatched DNA targets was studied effectively by differential pulse voltammetry. The proposed strategy could detect target DNA down to 3 × 10-11 M, with a linear range from 1 × 10-6 M to 1 × 10-10 M, showing high sensitivity. This electrochemical method is simple, free from indicator, and shows good selectivity. Hence, electrochemical biosensors are successfully demonstrated for the detection of K. pneumoniae.
Subject(s)
Biosensing Techniques/methods , Electrochemical Techniques/methods , Graphite/chemistry , Indoles/chemistry , Klebsiella pneumoniae/isolation & purification , Nanocomposites/chemistry , DNA Probes/chemistry , DNA, Single-Stranded/chemistry , Klebsiella pneumoniae/genetics , Nucleic Acid HybridizationABSTRACT
The assembly of quantum dots (QDs) in a simply method opens up opportunities to obtain access to the full potential of assembled QDs by virtue of the collective properties of the ensembles. In this study, quantum dots CdTe and graphene (Gr) nanocomposite was constructed for the simultaneous determination of uric acid (UA) and dopamine (DA). The CdTe QDs-Gr nanocomposite was prepared by ultrasonication and was characterized with microscopic techniques. The nanocomposite modified electrode was characterized by cyclicvoltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). Due to the synergistic effects between CdTe QDs and Gr, the fabricated electrode exhibited excellent electrochemical catalytic activities, good biological compatibility and high sensitivity toward the oxidation of UA and DA. Under optimum conditions, in the co-existence system the linear calibration plots for UA and DA were obtained over the range of 3-600 µM and 1-500 µM with detection limits of 1.0 µM and 0.33 µM. The fabricated biosensor also exhibits the excellent repeatability, reproducibility, storage stability along with acceptable selectivity.
Subject(s)
Biosensing Techniques/methods , Cadmium Compounds/chemistry , Dopamine/urine , Graphite/chemistry , Nanocomposites/chemistry , Quantum Dots , Tellurium/chemistry , Uric Acid/urine , Dielectric Spectroscopy , Electrodes , Humans , Limit of Detection , Oxidation-Reduction , Reproducibility of ResultsABSTRACT
To develop a new electrochemical DNA biosensor for determination of Klebsiella pneumoniae carbapenemase, a highly sensitive and selective electrochemical biosensor for DNA detection was constructed based on a glassy carbon electrode (GCE) modified with gold nanoparticles (Au-nano). The Au-nano/GCE was characterized by scanning electromicroscopy, cyclic voltammetry, and electrochemical impedance spectroscopy. The hybridization detection was measured by differential pulse voltammetry using methylene blue as the hybridization indicator. The dynamic range of detection of the sensor for the target DNA sequences was from 1 × 10(-11) to 1 × 10(-8) M, with an LOD of 1 × 10(-12) M. The DNA biosensor had excellent specificity for distinguishing complementary DNA sequence in the presence of non-complementary and mismatched DNA sequence. The Au-nano/GCE showed significant improvement in electrochemical characteristics, and this biosensor was successfully applied for determination of K. pneumoniae.
Subject(s)
Bacterial Proteins/analysis , Biosensing Techniques/methods , Klebsiella pneumoniae/enzymology , Metal Nanoparticles/chemistry , Oligonucleotides/genetics , beta-Lactamases/analysis , Carbon , Electrochemistry , Electrodes , Gold , Nucleic Acid HybridizationABSTRACT
We describe the fabrication of a sensitive electrochemical DNA biosensor for determination of Klebsiella pneumoniae carbapenemase (KPC). The highly sensitive and selective electrochemical biosensor for DNA detection was constructed based on a glassy carbon electrode (GCE) modified with gold nanoparticles (Au-NPs) and graphene (Gr). Then Au-NPs/Gr/GCE was characterized by scanning electro microscope (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The hybridization detection was measured by diffierential pulse voltammetry (DPV) using methylene blue (MB) as the hybridization indicator. The dynamic range of detection of the sensor for the target DNA sequences was from 1 × 10(-12) to 1 × 10(-7)mol/L, with a detection limit of 2 × 10(-13)mol/L. The DNA biosensor had excellent specificity for distinguishing complementary DNA sequence in the presence of non-complementary and mismatched DNA sequence. The results demonstrated that the Au-NPs/Gr nanocomposite was a promising substrate for the development of high-performance electrocatalysts for determination of KPC.