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For the successful treatment of infections, real-time analysis and enhanced multiplex capacity, sensitivity and cost-effectiveness of the developed detection method are critical. In this work, surface-enhanced Raman scattering (SERS) was employed with the final aim of identification and discrimination of pathogenic bacteria, based on their detected SERS fingerprint at the single-cell level. Several genera of bacteria that are found in most of the isolated infections in bacteraemia were successfully identified in less than 5 minutes without the use of antibodies or other specific receptors. The key element of the SERS direct detection platform is the SERS substrate, which combines easy production at low costs with a high enhancement enabling single-cell detection. The innovative approach of detection required the in situ synthesis of silver nanoparticles (NPs), ensuring an intimate contact with the bacterial membrane. This protocol provided a good reproducibility of the single-cell SERS spectra and was successfully applied both on Gram-negative and Gram-positive microorganisms (E. coli, M. morganii, E. lactis, L. casei). Thus, a label-free SERS-based biosensor for pathogen detection was developed with low costs, minimal sample preparation, high-accuracy and a very short analysis time of less than 5 min, which is crucial for infection diagnosis.

In this study, we aimed at developing a rapid and non‐destructive method to predict the genus‐level taxonomic affiliation of phenotypically similar halophilic archaea of Halobacteria Class. For this purpose, representatives of three widespread and frequently isolated haloarchaeal genera (Halobacterium sp., Haloferax sp. and Halorubrum sp.) were investigated by means of Raman spectroscopy. The most relevant Raman bands exhibited at 1505, 1150 and 1000 cm−1, respectively, were ascribed to the bacterioruberin carotenoid found in haloarchaea. Due to the similar spectral profiles, a robust chemometric approach based on fuzzy principal component analysis and linear discriminant analysis for the classification of the three halophilic strains was employed, and the high‐accuracy grouping of spectral data corresponding to those strains was achieved. Copyright © 2017 John Wiley & Sons, Ltd.

The world is in the midst of a pre-emptive public health emergency, one that is just as dramatic as the global aggressive viruses-related crises (Ebola, Zika, or SARS), but not as visible. The "superbugs" and their antimicrobial resistance do not cause much public alarm or awareness, but provoke financial losses of $100 trillion annually (WHO, http://www.who.int/mediacentre/commentaries/superbugs-action-now/en/ ). This status quo review offers an overview of ultrasensitive methods for high-throughput monitoring of bacteria during infection treatment, the effects of antibiotics on bacteria at single-cell level and the challenges we will face in their detection due to the extraordinary capability of these "superbugs" to gain and constantly improve multiresistance to antibiotics. A special emphasis is put on the ultrasensitive spectroscopic-based analysis techniques, using nanotechnology or not necessarily, that are more and more promising alternatives to conventional culture-based ones. The particular case of Mycobacteria detection is discussed based on recent reported work.

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