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Appl Environ Microbiol. 1972 April; 23(4): 714-720
Copyright © 1972 American Society for Microbiology. All Rights Reserved.

Separation of Treponema pallidum from Tissue Substances by Continuous-Flow Zonal Centrifugation

Myrtle L. Thomas, J. Walter Clark Jr., George B. Cline1, Norman G. Anderson2 and Harold Russell

a Venereal Disease Research Laboratory, Venereal Disease Branch, State and Community Services Division, Center for Disease Control, Atlanta, Georgia 30333

ABSTRACT

The zonal ultracentrifuge was used for separation of Treponema pallidum from large volumes of rabbit testicular syphiloma extracts by continuous-flow centrifugation in a cesium chloride density gradient. The gradient was linear with radius from a density of 1.05 to 1.36 g/ml. Operating speeds were 15,000 rev/min for the continuous-flow phase and 25,000 rev/min for a 30-min banding period. A total of 9 x 109 (24.3%) treponemes were recovered from the original extract. Of the treponemes recovered, 88% formed a band at a density of 1.170 to 1.211 g/ml. Within the limits of present methods of assay, these fractions were relatively free from testicular particulates and protein when compared with treponemes recovered after differential centrifugation. Observations of the isolated fractions by dark-field and electron microscopy indicated a lack of gross morphological damage to T. pallidum. Their antigenic characteristics were also retained, as evidenced by their ability to react with syphilitic sera in the indirect fluorescent-antibody procedure.

FOOTNOTES

1 University of Alabama, Birminghan, Ala. 36609.

2 Oak Ridge National Laboratory, Biology Division, Oak Ridge, Tenn.

Appl Environ Microbiol. 1972 April; 23(4): 714-720
Copyright © 1972 American Society for Microbiology. All Rights Reserved.





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Copyright © 1972 by the American Society for Microbiology. All rights reserved.