Sage Journals: Discover world-class research

Abstract

Mixtures containing two bacterial species were analyzed using flow cytometric techniques. Light scattering characteristics of Streptococcus mutans and Actinomyces viscosus show unique profiles for pure cultures. However, the light scatter analysis of a mixture containing these two species demonstrates overlapping near the origin. Thus, light scatter analysis was not sufficient to speciate bacteria with different morphologies. Labeling of samples with species-specific immunofluorescent antibodies permitted speciation of mixtures. As the percentage of the bacterium to which the antibody is directed increased in a two-component mixture, fluorescent flow cytometric analysis showed a corresponding increase in the percentage of cells displaying fluorescent labeling. These methods could permit the rapid identification of bacteria from oral sites without culturing.

Get full access to this article

View all access options for this article.

References

Arndt-Jovin, D.J. and Jovin, T.M. : Automated Cell Sorting with Flow Systems, Ann Rev Biophys Bioeng 7:527-558, 1978.

Buttke, T.M. ; Mallett, G.S. ; and Cuchens, M.A. : Positive Selection of Mouse B and T Lymphocytes and Analysis of Isolated Populations by Flow Cytometry, J Immunol Meth 58:193-207, 1983.

Darwish, S. ; Hyppa, T. ; and Socransky, S.S. : Studies of the Predominant Cultivable Microbiota of Early Periodontitis , J Periodont Res 13:1-16, 1978.

Emilson, C.G. ; Kohler, B. ; and Bratthall, D. : Immunofluorescent Determination of the Relative Proportions of Streptococcus mutans in Human Dental Plaque: A Comparison with Cultural Technique, Arch Oral Biol 20:81-86, 1974.

Evian, C.I. ; Rosenberg, E.S. ; and Listgarten, M.A. ; Bacterial Variability Within Diseased Periodontal Sites , J Periodontol 53:595-598, 1982.

Fazel-Madjlessi, J. ; Bailey, J.E. ; and McQuitty, D.N. : Flow Microfluorometry Measurements of Multicomponent Cell Composition During Batch Bacterial Growth, Biotech and Bioeng 22:457-462,1980.

Grenier, E.M. ; Eveland, W.C. ; and Loesche, W.J. : Identification of Streptococcus mutans Serotypes in Dental Plaque by Fluorescent Antibody Techniques, Arch Oral Biol 18:707-715, 1973.

Herzenberg, L.A. ; Sweet, R.G. ; and Herzenberg, L.A. : Fluorescence-Activated Cell Sorting, Sci Am 234:108-117, 1976.

Hutter, K.J. and Eipel, H.E. : Microbial Determinations by Flow Cytometry, J Gen Microbiol 113:369-375, 1979.

10.

Listgarten, M.A. and Hellden, L. : Relative Distribution of Bacteria at Clinically Healthy and Periodontally Diseased Sites in Humans, J Clin Periodontod 5:115-132, 1978.

11.

Newman, M.G. and Socransky, S.S. : Predominant Cultivable Microbiota in Periodontosis, J Periodont Res 12:120-128, 1977.

12.

Paau, A.S. ; Cowles, J.R. ; and Oro, J. : Flow-Microfluorometric Analysis of Escherichia coli, Rhizobium meliloti and Rhizobium japonicum at Different Stages of the Growth Cycle, Can J Microbiol 23:1165-1169, 1977.

13.

Proceedings of Workshop on Quantitative Evaluation of Periodontal Diseases by Physical Measurement Techniques, J Dent Res 58:547-553,1979.

14.

Slots, J. : Subgingival Microflora and Periodontal Disease, J Clin Periodontol 6:351-382, 1979.

15.

Smulow, J.B. ; Turesky, S.S. ; and Hill, R.G. : The Effect of Supragingival Plaque Removal on Anaerobic Bacteria in Deep Periodontal Pockets, JADA 107:737-742, 1983.

16.

Socransky, S.S. ; Gibbons, R.J. ; Dale, A.C. ; Bortnick, L. ; Rosenthal, E. ; and MacDonald, J.B. : The Microbiota of the Gingival Crevice Area of Man, Arch Oral Biol 8:275-280, 1963.

17.

Socransky, S.S. : Microbiology of Periodontal Disease - Present Status and Future Considerations , J Periodontol 48:497-504, 1977.

18.

Thomson, L.A. ; Little, W. ; and Hegeagv, G.J. : Application of Fluorescent Antibody Methods in Analysis of Plaque Samples , J Dent Res 55(A):80-86, 1976.

19.

Thomson, L.A. : Preparation of Antiserums for Use in the Fluorescent Antibody Identification of Certain Plaque Bacteria, J Dent Res 55(A):28-32, 1976.

20.

Van Dilla, V.S. : Langlois, R.G. ; Pinkel, D. ; Yajko, D. ; and Hadley, W.K. : Bacterial Characterization by Flow Cytometry, Science 220:620-622, 1983.

21.

Wilson, M.W. : The Agar Diffusion Precipitation Technique. A Comparison of the Simple and Double Diffusion, J Immunol 81:317-330, 1958.

Automated Immunofluorescent Speciation of Oral Bacteria Using Flow Cytometry

Abstract

Get full access to this article

References