研究目的
To develop a rapid SERS-based method for the identification of methicillin-resistant Staphylococcus aureus (MRSA) using positively charged silver nanoparticles, aiming to provide a quick, reproducible, and accurate diagnostic tool for bacterial infections and antibiotic resistance.
研究成果
The SERS method using AgNPs+ successfully identified MRSA with high accuracy (almost 100%) in a rapid assay time of 45 minutes. It differentiated S. aureus from other microorganisms and various antibiotic-resistant strains, demonstrating potential as a low-cost tool for rapid detection of antibiotic resistance. Future work should focus on reducing detection time, clarifying molecular mechanisms, and expanding the database for broader applications.
研究不足
The method requires bacterial isolation culture, taking almost 25 hours total time including culture and SERS test, which is not as fast as desired for clinical use. The molecular mechanism of SERS spectral differences between MSSA and MRSA is unclear, and the assignment of the 1523 cm?1 Raman band needs further study. The number of clinical samples is insufficient for a standard discriminant analysis model.
1:Experimental Design and Method Selection:
The study employed a SERS-based approach using positively charged silver nanoparticles (AgNPs+) synthesized with CTAB as a stabilizer. The method relies on electrostatic self-assembly of AgNPs+ on bacterial surfaces to enhance Raman signals for pathogen identification and discrimination.
2:Sample Selection and Data Sources:
Six standard bacterial strains (S. aureus 29213, S. aureus 25923, C. albicans, B. cereus, E. coli, P. aeruginosa) and clinical isolates (52 MSSA and 215 MRSA) were used. Strains were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China) and provided by the Clinical Microbiology Laboratory, Southwest Hospital.
3:List of Experimental Equipment and Materials:
Equipment included a scanning electron microscope (Crossbeam 340, Zeiss Inc.), Zetasizer analyzer (Zetasizer Nano ZSP, Malvern Inc.), spectrophotometer (NanoDrop 2000, Thermo Scientific Inc.), Raman micro-spectrometer (XploRA PLUS, Horiba Scientifics), bacteria counter (THOMA AUVON), and automatic microbial identification system (Vitek?2 Compact system, biomerieux). Materials included AgNO3, NaBH4, CTAB, and other chemicals from Sangon Biotechnology Inc., and ultrapure water from Milli-Q System.
4:Experimental Procedures and Operational Workflow:
AgNPs+ were synthesized and characterized. Bacterial samples were cultured, prepared to OD600=
5:0, mixed with AgNPs+ at a
1 volume ratio, incubated for 30 minutes, pipetted onto Si chips, dried, and SERS spectra were collected using a 532 nm laser with specific parameters (e.g., 10 s exposure time).
6:Data Analysis Methods:
SERS spectra were processed using LabSpec 6 software for smoothing, baseline correction, and normalization. Statistical analyses (PLS-DA and OPLS-DA) were performed with SIMCA 14.0 software, including cross-validation to assess model performance.
独家科研数据包,助您复现前沿成果,加速创新突破
获取完整内容