Citation Index 6
Multispectral Bacterial Identification
Michael A. Tanner, William J. Coleman, Christine L. Everett, Steven J. Robles,
Michael R. Dilworth, Mary M. Yang, and Douglas C. Youvan
Bldg., 62*
3350 Scott Blvd.
Santa Clara, CA 95054 USA
Abstract
A multispectral optical technique was developed to simultaneously classify individual bacterial cells within mixed populations. Multispectral Bacterial Identification (mBID) combines innovations in microscopy with a software analysis program to measure and categorize the fluorescence signals from multiplexed 16S ribosomal RNA probes hybridized to populations of different bacteria. Software was developed to identify individual bacteria at the level of species within these mixed populations. To test the feasibility of mBID, we examined the fluorescence emissions from a mixture of probes specific for individual species of known bacteria from the American Type Culture Collection (ATCC). Currently, up to seven species can be detected simultaneously by fluorescence microscopy. An eighth signal was reserved for a universal probe to control for fluorescence intensity. mBID can also be used to identify uncultured microorganisms. We plan to couple this new multispectral technology to existing identification technologies that utilize 16S rRNA sequence alignment. Using this integrated identification protocol, bacteria that may be associated with chronic conditions (e.g., prostatitis and vaginosis) will be identified first by analyzing their 16S rDNA sequences and then by visualizing them with fluorescent probes hybridized to their 16S rRNA in situ.
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© Biotechnology et alia, 2000
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