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Surface-enhanced Raman scattering method for the identification of methicillin-resistant Staphylococcus aureus using positively charged silver nanoparticles

DOI:10.1007/s00604-018-3150-6 期刊:Microchimica Acta 出版年份:2019 更新时间:2025-09-23 15:22:29
摘要: The article describes a SERS-based method for diagnosis of bacterial infections. Positively charged silver nanoparticles (AgNPs+) were employed for identification of methicillin-resistant Staphylococcus aureus (MRSA). It is found that AgNPs+ undergo self-assembly on the surface of bacteria via electrostatic aggregation. The assembled AgNPs+ are excellent SERS substrates. To prove the capability of SERS to differentiate between S. aureus and other microorganisms, six standard strains including S. aureus 29213, S. aureus 25923, C. albicans, B. cereus, E. coli, and P. aeruginosa were tested. To further demonstrate its applicability for the identification of MRSA in clinical samples, 52 methicillin-sensitive S. aureus (MSSA) isolates and 215 MRSA isolates were detected by SERS. The total measurement time (include incubation) is 45 min when using a 3 μL sample. The method gives a strongly enhanced Raman signal (at 730 cm?1 and 1325 cm?1) with good reproducibility and repeatability. It was successfully applied to the discrimination of the six strain microorganisms. The typical Raman peaks of S. aureus at 730, 1154, 1325, and 1457 cm?1 were observed, which were assigned to the bacterial cell wall components (730 cm?1- adenine, glycosidic ring mode, 1154 cm?1- unsaturated fatty acid, 1325 cm?1- adenine, polyadenine, and 1457 cm?1 for -COO- stretching). S. aureus was completely separated from other species by partial least squares discriminant analysis (PLS-DA). Moreover, 52 MSSA isolates and 215 MRSA isolates from clinical samples were identified by PLS-DA. The accuracy was almost 100% when compared to the standard broth microdilution method. A classification based on latent structure discriminant analysis provided spectral variability directly. Conceivably, the method offers a potent tool for the identification of bacteria and antibiotics resistance, and for studies on antibiotic-resistance in general.
作者: Xueping Chen,Meiqiong Tang,Yu Liu,Jiaoqi Huang,Zhiyong Liu,Huiyan Tian,Yuting Zheng,Marc Lamy de la Chapelle,Yang Zhang,Weiling Fu
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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.

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.

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.

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