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Rapid method for SV40 titration.

Nir Drayman1, Slava Kler, Orly Ben-nun-Shaul

  • 1Department of Hematology, Hebrew University-Hadassah Medical School, Ein Kerem, Jerusalem, Israel.

Journal of Virological Methods
|December 17, 2009
PubMed
Summary
This summary is machine-generated.

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A new flow cytometry assay simplifies Simian virus 40 (SV40) titration, providing results in two days instead of two weeks. This method also revealed variations in SV40 susceptibility across different monkey kidney cell lines.

Area of Science:

  • Virology
  • Cell Biology
  • Immunology

Background:

  • Traditional Simian virus 40 (SV40) titration relies on plaque assays, which are time-consuming (approximately two weeks).
  • This delay can impede research progress in virology and related fields.
  • A faster, more efficient method for determining viral titers is needed.

Purpose of the Study:

  • To develop and validate a simplified assay for SV40 titration.
  • To reduce the time required for SV40 titer determination.
  • To investigate potential differences in SV40 susceptibility among various cell lines.

Main Methods:

  • A flow cytometry-based assay was developed utilizing T-antigen detection.
  • Cells were infected with serial dilutions of SV40, harvested 48 hours post-infection, and stained for T-antigen.

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  • Viral titer was calculated based on the percentage of T-antigen positive cells.

    • Main Results:

    • The new assay provides SV40 titers within two days, significantly faster than conventional plaque assays.
    • Consistent differences in SV40 titers were observed across different permissive African Green Monkey kidney cell lines.
    • This suggests variable susceptibility of these cell lines to SV40 infection.

    Conclusions:

    • The flow cytometry assay is a rapid and efficient method for SV40 titration.
    • The findings highlight the importance of cell line selection for SV40 studies due to variable susceptibility.
    • This optimized method is readily adaptable for titrating other viruses.