We have investigated the fluorescence profiles of four common pathogens: S. pneumoniae, S. aureus, M. catarrhalis, and H. influenzae. The steady-state auto fluorescence spectra of bacteria are measured as a function of the incident light from 200 to 700 nm. The spectra for each bacterium are combined into a fluorescence profile or fluorescence finger print. Each bacterium produces a unique in vitro fluorescence profile when measured in a saline suspension. The profiles are reproducible. Suspensions of a bacterial strain, where the identification is not known, can be correctly matched to a small library of previously measured fluorescence profiles using a linear least-squares fitting algorithm. In addition, we have measured the fluorescence and absorption spectrum of the tympanic membrane removed from a chinchilla. The optical properties of the tympanic membrane and the least-squares identification process form precept for a non-invasive, fluorescence based bacterial diagnosis technique to be used in otitis media.
Currently, the identification of the bacteria responsible for acute otitis media requires a painful invasive procedure: tympanocentesis. To develop a rapid and noninvasive technique for bacterial diagnosis, the fluorescence profiles of four common pathogens and the optical characteristics of the tympanic membrane have been investigated. Each bacterium produces a unique in vitro fluorescence profile when measured in a saline suspension. Also, spectrally resolved transmission measurements from the chinchilla tympanic membrane demonstrate an optical window that will transmit sufficient light for in vivo measurement of the fluorescence profiles. Thus, we have established the precept for a fluorescence-based bacterial diagnosis technique to be used in otitis media. This paper presents the theory, optical data, and a discussion of the device engineering involved in the technique.
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