In situ detection of live-to-dead bacteria ratio after inactivation by means of synchronous fluorescence and PCA

Significance The in situ and rapid detection of live and dead bacteria is crucial for preventing health care-associated infection in hospitals, field clinics, and other areas such as monitoring wound healings. Traditionally, determining the number of pathogenic organism involves specimen collection, appropriate culture on agar plates within laboratory environments, and counting the colony-forming units. These are very time-consuming procedures and could therefore increase the risk of infection or even death. In this paper, we demonstrate that the live/dead bacteria ratio after UV radiation and antibiotic treatment may be obtained easily, through a handheld spectrophotometer integrated with principal component analysis. Such procedures may provide a new way for the rapid detection and classification of live and dead bacteria in situ within minutes. The determination of live and dead bacteria is of considerable significance for preventing health care-associated infection in hospitals, field clinics, and other areas. In this study, the viable (live) and nonviable (dead) bacteria in a sample were determined by means of their fluorescence spectra and principal component analysis (PCA). Data obtained in this study show that it is possible to identify bacteria strains and determine the live/dead ratio after UV light inactivation and antibiotic treatment, in situ, within minutes. In addition, synchronous fluorescence scans enable the identification of bacterial components such as tryptophan, tyrosine, and DNA. Compared with the time-consuming plating and culturing methods, this study renders a means for rapid detection and determination of live and dead bacteria.

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