ABSTRACT
A multifunctional surface-enhanced Raman scattering (SERS) platform integrating sensitive detection and drug resistance analysis was developed for Gram-positive bacteria. The substrate was based on self-assembled Ti3C2Tx@Au NPs films and capture molecule phytic acid (IP6) to achieve specific capture of Gram-positive bacteria and different bacteria were analyzed by fingerprint signal. It had advantages of good stability and homogeneity (RSD = 8.88%). The detection limit (LOD) was 102 CFU/mL for Staphylococcus aureus and 103 CFU/mL for MRSA, respectively. A sandwich structure was formed on the capture substrate by signal labels prepared by antibiotics (penicillin G and vancomycin) and non-interference SERS probe molecules (4-mercaptobenzonitrile (2223 cm-1) and 2-amino-4-cyanopyridine (2240 cm-1)) to improve sensitivity. The LOD of Au NPs@4-MBN@PG to S. aureus and Au NPs@AMCP@Van to MRSA and S. aureus were all improved to 10 CFU/mL, with a wide dynamic linear range from 108 to 10 CFU/mL (R2 ≥ 0.992). The SERS platform can analyze the drug resistance of drug-resistant bacteria. Au NPs@4-MBN@PG was added to the substrate and captured MRSA to compare the SERS spectra of 4-MBN. The intensity inhomogeneity of 4-MBN at the same concentrations of MRSA and the nonlinearity at the different concentrations of MRSA revealed that MRSA was resistant to PG. Finally, the SERS platform achieved the determination of MRSA in blood. Therefore, this SERS platform has great significance for the determination and analysis of Gram-positive bacteria.
Subject(s)
Anti-Bacterial Agents , Gold , Limit of Detection , Metal Nanoparticles , Spectrum Analysis, Raman , Staphylococcus aureus , Titanium , Spectrum Analysis, Raman/methods , Gold/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Titanium/chemistry , Metal Nanoparticles/chemistry , Staphylococcus aureus/drug effects , Staphylococcus aureus/isolation & purification , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Vancomycin/pharmacology , Vancomycin/chemistry , Drug Resistance, Bacterial , Microbial Sensitivity Tests , Penicillin G/pharmacology , Penicillin G/chemistry , Gram-Positive Bacteria/drug effects , Gram-Positive Bacteria/isolation & purificationABSTRACT
The harm and impact of the COVID-19 pandemic have highlighted the importance of fast, sensitive, and cost-effective virus detection methods. In this study, we developed a DNA aptamer sensor using nanoparticle-enhanced surface plasmon resonance imaging (SPRi) technology to achieve efficient labeling-free detection of SARS-CoV-2 S protein. We used the same DNA aptamer to modify the surface of the SPRi sensor chip and gold nanoparticles (AuNPs), respectively, for capturing target analytes and amplifying signals, achieving ideal results while greatly reducing costs and simplifying the preparation process. The SPRi sensing method exhibits a good linear relationship (R2 = 0.9926) in the concentration range of 1-20 nM before adding AuNPs to amplify the signal, with a limit of detection (LOD) of 0.32 nM. After amplifying the signal, there is a good linear relationship (R2 = 0.9829) between the concentration range of 25-1000 pM, with a LOD of 5.99 pM. The simulation results also verified the effectiveness of AuNPs in improving SPRi signal response. The SPRi sensor has the advantage of short detection time and can complete the detection within 10 min. In addition, the specificity and repeatability of this method can achieve excellent results. This is the first study to simultaneously capture a viral marker protein and amplify the signal using polyadenylic acid (polyA)-modified DNA aptamers on the SPR platform. This scheme can be used as a fast and inexpensive detection method for diagnosis at the point of care (POC) to combat current and future epidemics caused by the virus.
Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , COVID-19 , Metal Nanoparticles , Humans , Surface Plasmon Resonance/methods , Spike Glycoprotein, Coronavirus , Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , Gold/chemistry , Pandemics , Metal Nanoparticles/chemistry , COVID-19/diagnosis , SARS-CoV-2 , DNA , Viral ProteinsABSTRACT
Recent years, two-dimensional transition metal carbonitrides (MXene) have attracted much attention in the field of surface-enhanced Raman scattering (SERS). However, the relatively low enhancement of MXene is a major challenge. Herein, Nb2C-Au NPs nanocomposites were prepared by electrostatic self-assembly method, which have a synergistically conjugated SERS effect. The EM hot spots of Nb2C-Au NPs are significantly enlarged and expanded, while the surface Fermi level is decreased. This synergistic effect could improve the SERS performance of the system. Consequently, for the dye molecules CV and MeB, the detection limits reach 10-10 M and 10-9 M, respectively, while for biomolecule adenine, the detection limit is as low as 5 × 10-8 M. The results also show the good concentration-dependent linearity, uniformity, reproducibility and stability of SERS substrate. Nb2C-Au NPs could be a fast, sensitive and stable SERS platform for label-free and non-destructive detection. This work may expand the application of MXene based materials in the field of SERS.
