Noninvasive optical diagnosis of bacteria causing otitis media

Werkhaven, Jay A.; Reinisch, Lou; Sorrell, M.; Tribble, Jerri A.; Ossoff, Robert H.

doi:10.1288/00005537-199403000-00004

Cited by 22 publications

(8 citation statements)

References 14 publications

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“…First, we (Pinnick et al, 1995) and others (Hairston et al, 1997) have shown that fluorescence particle counters that measure the elastic scattering and total (undisperscd) fluorescence can differentiate between some bioloeical " and nonbiological aerosols; the advantage of obtaining single-particle measurements (as opposed to volumc scattering measurements on many particles) are cited above. Second, excitation/emission spectra of the intrinsic fluorescence of biological particles have been shown to be useful for differentiating between a few types of biological (Sorrel1 et al, 1994;Werkhaven et al, 1994). These spectra and similar more extensive excitation/emission spectra measured for a number of microorganisms in liquid suspension (Steve Gotoff, private communication March 1997) suggest that excitation with multiple wavelengths will be necessary for more-than a mostrudimentary classification.…”

Section: Introductionmentioning

confidence: 96%

Aerosol Fluorescence Spectrum Analyzer for Rapid Measurement of Single Micrometer-Sized Airborne Biological Particles

Pinnick

Hill

Nachman

et al. 1998

Aerosol Science and Technology

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We report the operation of an aerosol analyzer capable of measuring the fluorescence spectra of single micrometer-sized bioaerosol particles as they flow through the instrument. Particles entrained in an airstream initially traverse a cw (continuous wave) (488 nm) "trigger" laser beam where their elastic scattering and total fluorescence is measured with photomultipliers. When the elastic scattering andlor fluorescence signals exceed preset levels, a 266 nm UV "probe" laser is triggered to fire and illuminate preselected particles downstream from the trigger laser. The UV laser-excited spectra of particles are measured with the instrument's image-intensified CCD detector that is gated to be on when the probe laser fires. We demonstrate the ability of the instrument to capture the fluorescence spectra of single micrometer-sized airborne biological particles. Such spectra should be useful in differentiating between biological and nonbiological aerosols and in partially characterizing airborne biological particles.

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Section: Introductionmentioning

confidence: 96%

Aerosol Fluorescence Spectrum Analyzer for Rapid Measurement of Single Micrometer-Sized Airborne Biological Particles

Pinnick

Hill

Nachman

et al. 1998

Aerosol Science and Technology

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“…Werkhaven et al 2 acknowledged the potential for the tympanic membrane to create its own fluorescence signal and absorb a portion of the excitation and emission wavelengths in vivo. Thus they measured the emission pattern and absorption spectrum of a tympanic membrane from a healthy animal in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…Previous research by Werkhaven et al 2 and Sorrell et al 3 has established the groundwork for accurate identification of common bacterial pathogens using fluorescence emission spectroscopy. In their investigation, S pneumoniae , H influenzae , M catarrhalis , and Staphylococcus aureus were each suspended in saline, placed in a quartz cuvette, and successively differentiated in vitro by analysis of their emission spectra.…”

Section: Introductionmentioning

confidence: 99%

Noninvasive Fluorescent Identification of Bacteria Causing Acute Otitis Media in a Chinchilla Model

Spector¹,

Reinisch²,

Smith³

et al. 2000

The Laryngoscope

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Objectives: To evaluate a noninvasive method of bacterial identification via fluorescence spectroscopy in the setting of acute otitis media in a chinchilla model. Study Design: Prospective. Methods: For each chinchilla, transbullar inoculation with Streptococcus pneumoniae, Haemophilus influenzae, or Staphylococcus aureus was performed bilaterally and clinical infection was determined by otoscopy. An optical fiber coupled to a spectrofluorometer allowed for the delivery of an excitation wavelength to the inflamed tympanic membrane and the acquisition of the resulting emission signal. Sequential emission spectra obtained over a range of excitation wavelengths were assembled by a computer algorithm, and a single, three-dimensional plot was created for each test ear. Similarly, plots from the healthy external auditory canal (EAC) were also recorded. Twelve animals were used to establish a library of four reference plots representing the three bacteria and the EAC. Of the 24 ears available for study, 10 were excluded from analysis because of lack of clinical infection or presence of tympanic membrane perforation with purulent drainage. From four additional animals, four samples from ears infected with the above bacteria and three samples of the EAC served as unknowns. The unknown plots were analyzed by an investigator blinded to their identity. Results: Using a multiple correlation of the unknown to the reference plots, seven of seven samples were correctly identified. Conclusions: We were able to establish a prototype method for the noninvasive identification of a limited library of pathogens in a chinchilla model of acute otitis media. Key Words: Acute otitis media, fluorescence, noninvasive diagnosis, chinchilla.Laryngoscope , 110:1119-1123, 2000 . INTRODUCTIONAppropriate man agement of acute otitis media (AOM) requires an under standing of bact eria and their propensity to alter sensitivities to antimicrobial th era py. Important vari ables to consider include the prevalence, n atural history, and r esistance patterns of common pathogens ca using AOM within each community. 1 The presence of resistant bacteri a should be su spected in the setting of persistent clinical infec tion despite an initial CO\.ll"Se of treatment. Currently the most common pa thogens that limit the efficacy of first-line ther apy include mul tidr ugr esistant Streptococcus pneumoniae a nd {3-lactamase-producing strains of Haemophilus influenzae and Moraxella catarrhalis. With this in mind , physici ans may tailor th eir n ext treatment selection to best counter known mech anisms of antibiotic resistance. Examples include incr easing the dose of amoxicillin to heighten the concentration of drug in the middle ear fluid or selecting a second-line antibiotic with better activity against {3-lactamase-producing species and penicillin-resista nt S pneumoniae.At present, bacterial identifica tion in the setting of AOM can be accomplished using needl e aspiration of the middle ear fluid via tympanocentesis. Di sadvantages of this procedure inc...

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“…Native fluorescence from bacteria in the visible and UV is dominated by tryptophan emission. [1][2][3][4][5][6] Emission in the 430-to 514-nm region from pterins, nicotinamide adenine dinucleotide, nicotinamide adenine dinucleotide phosphate, and flavin adenine dinucleotides have also been observed. 3 The development of bacterial endospores by Bacillus subtilis ͑Bs͒ represents one of the best-studied models for the observation of the well-defined process of spore formation.…”

Section: Introductionmentioning

confidence: 94%

Native fluorescence and excitation spectroscopic changes in Bacillus subtilis and Staphylococcus aureus bacteria subjected to conditions of starvation

Alimova

Katz²,

Savage³

et al. 2003

Appl. Opt.

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Fluorescence emission and excitation spectra were measured over a 7-day period for Bacillus subtilis (Bs), a spore-forming, and Staphylococcus aureus (Sa), a nonspore-forming bacteria subjected to conditions of starvation. Initially, the Bs fluorescence was predominantly due to the amino acid tryptophan. Later, a fluorescence band with an emission peak at 410 nm and excitation peak at 345 m, from dipicolinic acid, appeared. Dipicolinic acid is produced during spore formation and serves as a spectral signature for detection of spores. The intensity of the 410-nm band continued to increase over the next 3 days. The Sa fluorescence was predominantly from tryptophan and did not change over time. In 6 of the 17 Bs specimens studied, an additional band appeared with a weak emission peak at 460 cm and excitation peaks at 250, 270, and 400 nm. The addition of beta-hydroxybutyric acid to the Bs or the Sa cultures resulted in a two-order of magnitude increase in the 460-nm emission. The addition of Fe2+ quenched the 460 emission, indicating that a source of the 460-nm emission was a siderophore produced by the bacteria. We demonstrate that optical spectroscopy-based instrumentation can detect bacterial spores in real time.

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Noninvasive optical diagnosis of bacteria causing otitis media

Cited by 22 publications

References 14 publications

Aerosol Fluorescence Spectrum Analyzer for Rapid Measurement of Single Micrometer-Sized Airborne Biological Particles

Aerosol Fluorescence Spectrum Analyzer for Rapid Measurement of Single Micrometer-Sized Airborne Biological Particles

Noninvasive Fluorescent Identification of Bacteria Causing Acute Otitis Media in a Chinchilla Model

Native fluorescence and excitation spectroscopic changes in Bacillus subtilis and Staphylococcus aureus bacteria subjected to conditions of starvation

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