1887

Abstract

SUMMARY

A thin sectioning procedure was used to detect and enumerate oncornavirus particles in tumour-cell culture fluids, tumour homogenates, mouse blood plasma, mouse and human milk specimens, and density gradient fractions of these specimens. Oncornaviruses studied included mouse mammary tumour, murine sarcoma, ESP-1, and RD-114 viruses. In this procedure particles were sedimented on to small membrane filter discs in the ultracentrifuge using inexpensive commercially available adapters and tubes. Particles in cross sections of the disc were counted and their total number determined by relating the effective surface area of a field to the surface area of the entire membrane disc. Particles may be reliably identified and counted in preparations containing predominately cellular debris. Linear dose response plots were obtained in serial dilution experiments utilising vaccinia virus, adenovirus type 2, and murine sarcoma virus, demonstrating the reliability of the procedure and its wide applicability. The counts obtained for adenovirus and vaccinia virus preparations were comparable to counts obtained for the same preparations in other laboratories by established methods. Statistically reliable counts have been obtained using sample vol. of 0.01 ml or less.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-21-1-57
1973-10-01
2022-01-22
Loading full text...

Full text loading...

/deliver/fulltext/jgv/21/1/JV0210010057.html?itemId=/content/journal/jgv/10.1099/0022-1317-21-1-57&mimeType=html&fmt=ahah

References

  1. Calafat J., Hageman P. C. 1973; Remarks on virus-like particles in human breast cancer. Nature, London 242:260–262
    [Google Scholar]
  2. Dmochowski L., Seman G., Myers B., Gallagher S. H. 1968; Relationship of viruses to the origin of human breast cancer. Medical Record and Annals 61:384–387
    [Google Scholar]
  3. Feller W. F., Chopra H. C. 1971; Virus-like particles in human milk. Cancer 28:1425–1430
    [Google Scholar]
  4. Friedman B., Blais P., Shaffer P. 1968; Fine structure of millipore filters. Journal of Cell Biology 39:208–211
    [Google Scholar]
  5. Gehle W. D., Smith K. O. 1970; Enumeration of virus particles in ultrathin sectioned pellets. Proceedings of the Society for Experimental Biology& Medicine 135:490–494
    [Google Scholar]
  6. Lyons M. J., Moore D. H. 1965; Isolation of the mouse mammary tumor virus: Chemical and morphological studies. Journal of the National Cancer Institute 35:549–565
    [Google Scholar]
  7. McAllister R. M., Nicholson N., Gardner M. B., Rongey R. W., Rasheed S., Sarma P. S., Huebner R. J., Hatanaka M., Oroszlan S., Gilden R. V., Kabigting A., Vernon L. 1972; C-type virus released from cultured human rhabdomyosarcoma cells. Nature New Biology 235:3–6
    [Google Scholar]
  8. McCombs R. M., Benyesh-Melnick M., Brunschwig J. P. 1968; The use of Millipore filters in ultrastructure studies of cell cultures and viruses. Journal of Cell Biology 36:231–243
    [Google Scholar]
  9. Miller M. F., Allen P. T., Bowen J. M., Dmochowski L., Hixson D. C., Williams W. C. 1972; Particle counting of partially purified RNA tumor viruses by a thin sectioning technique. Proceedings of 30th Annual Meeting Electron Microscopy Society of America pp 292–293 Edited by Arceneaux C. J. Baton Rouge, La., U.S.A.: Claitor’s Publishing Division;
    [Google Scholar]
  10. Peachey L. D. 1958; Thin sections. I. A study of section thickness and physical distortion produced during microtomy. Journal of Biophysical and Biochemical Cytology 4:233–242
    [Google Scholar]
  11. Priori E. S., Dmochowski L., Myers B., Wilbur J. R. 1971; Constant production of type C virus particles in a continuous tissue culture derived from pleural effusion cells of a lymphoma patient. Nature New Biology 232:61–62
    [Google Scholar]
  12. Sarkar N. H., Moore D. H. 1972; On the possibility of a human breast cancer virus. Nature, London 236:103–106
    [Google Scholar]
  13. Sharp D. G. 1949; Ennumeration of virus particles by electron micrography. Proceedings of the Society for Experimental Biology and Medicine 70:54–59
    [Google Scholar]
  14. Sharp D. G. 1965; Quantitative use of the electron microscope in virus research. In Quantitative Electron Microscopy pp 93–125 Edited by Bahr G. F, Zeitler E. H. Baltimore: Williams and Wilkins;
    [Google Scholar]
  15. Sharp D. G., Beard J. W. 1952; Counts of virus particles by sedimentation on agar and electron micrography. Proceedings of the Society for Experimental Biology and Medicine 81:75–79
    [Google Scholar]
  16. Smith K. O., Melnick J. L. 1962; A method for staining virus particles and identifying their nucleic acid type in the electron microscope. Virology 17:480–490
    [Google Scholar]
  17. Soehner R. L., Dmochowski L. 1969; Induction of bone tumors in rats and hamsters with murine sarcoma virus and their cell-free transmission. Nature, London 224:191–192
    [Google Scholar]
  18. Watson D. H. 1962; Electron-micrographic particle counts of phosphotungstate-sprayed virus. Biochimica et biophysica acta 61:321–331
    [Google Scholar]
  19. Williams R. C., Backus R. C. 1949; Macromolecular weights determined by direct particle counting. I. The weight of the bushy stunt virus particle. Journal of the American Chemical Society 71:4052–4057
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-21-1-57
Loading
/content/journal/jgv/10.1099/0022-1317-21-1-57
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error