Surface Enhanced Raman Spectroscopy for the Rapid Detection and Identification of Microbial Pathogens in Human Serum
Abstract
A rapid and sensitive means to diagnose bacterial agents would decrease the time to diagnosis, allow providers to administer the most effective antimicrobial earlier during the course of treatment and reduce mortality and morbidity due to infection. Biosensors based on Surface Enhanced Raman Spectroscopy (SERS) hold great promise for the rapid and sensitive detection of bacterial pathogens of military interest. The objective of this study was to evaluate a hand-held SERS-based diagnostic system to identify bacteria in pooled human sera using lysis filtration as a means of separating bacteria. Furthermore, the usefulness of pure culture bacteria in generating a reference library for identification of bacterial species recovered from serum was examined. Measurement of SERS spectra was carried out for bacteria from pure culture and bacteria recovered from pooled human sera using lysis filtration. Acenitobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa were evaluated for generation of SERSbased molecular fingerprints followed by Principal Component Analysis and Partial Least Squares Differential Analysis to determine uniqueness and commonalities of measured spectra. The successful detection, identification and classification of bacteria from human serum using a hand-held Raman spectrometer were demonstrated. Pure culture bacteria were readily identifiable and distinguishable by their SERS-based molecular fingerprints at the species level, but greater similarities existed in SERS spectra amongst different species recovered from serum. Lysis filtration successfully recovered hydrophilic bacteria without significantly affecting SERS spectra. However, shifts in relative peak intensities of SERS spectra were observed primarily for hydrophobic bacteria after recovery from serum. Hydrophilic bacteria, such as A. baumannii and S. aureus, recovered from serum using lysis filtration can be rea
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 11, 2014
- Accession Number
- ADA620292
Entities
People
- Christian N. Kotanen
- Luis A. Martinez
- Rene Alvarez
Organizations
- Naval Medical Research Center