Skip to main content

Detection of SV40 DNA Sequences in Human Tissue

  • Protocol
SV40 Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 165))

Abstract

Several recent investigations using polymerase chain reaction (PCR) detection methods have identified SV40-like DNA sequences in human neoplasms, particularly choroid plexus tumors (1), ependymomas (1), mesotheliomas (2), and osteosarcomas (3,4). Difficulties have arisen because of the use of paraffin-fixed tissue, loss of possible viral episomes during DNA isolation, low copy numbers of viral DNA, PCR contamination, and confusion with BKV and JCV sequences. Nonetheless, the authentication of amplified products by DNA sequencing, the finding of a single 72-bp viral-enhancer repeat element, variability of large tumor antigen (T-ag) carboxy terminal DNA sequences in many specimens, and the isolation of SV40 virus from a choroid plexus tumor all support the validity of the basic observation that SV40 is present in humans, and may therefore contribute to oncogenesis in special situations.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Bergsagel, D. J., Finegold, M. J., Butel, J. S., Kupsky, W. J., and Garcea, R. L. (1992) DNA sequences similar to those of simian virus 40 in ependymomas and choroid plexus tumors of childhood. N. Engl. J. Med. 326, 988–993.

    Article  CAS  PubMed  Google Scholar 

  2. Carbone, M., Pass, H. I., Rizzo, P., Marinetti, M., Di Muzio, M., New, D. J., et al. (1994) Simian virus 40-like DNA sequences in human pleural mesothelioma. Oncogene 9, 1781–1790.

    CAS  PubMed  Google Scholar 

  3. Carbone, M., Rizzo, P., Procopio, A., Giuliano, M., Pass, H. I., Gebhardt, M. C., et al. (1996) SV40-like sequences in human bone tumors. Oncogene 13, 527–535.

    CAS  PubMed  Google Scholar 

  4. Lednicky, J. A., Stewart, A. R., Jenkins, J. J. I., Finegold, M. J., and Butel, J. S. (1997) SV40 DNA in human osteosarcomas shows sequence variation among T-antigen genes. Int. J. Cancer 72, 791–800.

    Article  CAS  PubMed  Google Scholar 

  5. Impraim, C. C., Saiki, R. K., Erlich, H. A., and Teplitz, R. L. (1987) Analysis of DNA extracted from formalin-fixed, paraffin-embedded tissues by enzymatic amplification and hybridization with sequence-specific oliogonucleotides. Biochem. Biophys. Res. Commun. 142, 710–716.

    Article  CAS  PubMed  Google Scholar 

  6. Kwok, S. and Higuchi, R. (1989) Avoiding false positives with PCR. Nature (London) 339, 237,238.

    Article  CAS  Google Scholar 

  7. Lednicky, J. A. and Butel, J. S. (1997) A coupled PCR and restriction digest method for the detection and analysis of the SV40 regulatory region in infectedcell lysates and clinical samples. J. Virol. Methods 64, 1–9.

    Article  CAS  PubMed  Google Scholar 

  8. Butel, J. S. Arrington, A. S., Wong, C., Lednicky, J. A., and Finegold, M. J. (1999) Molecular evidence of SV40 infections in children. J. Infect. Dis. 180, 884–887.

    Article  CAS  PubMed  Google Scholar 

  9. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  10. Greer, C. E., Wheeler, C. M., and Manos, M. M. (1995) PCR amplification from paraffin-embedded tissues: sample preparation and the effects of fixation, in PCR Primer: A Laboratory Manual (Dieffenbach, C. W. and Dveksler, G. S., eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  11. Strickler, H. D., Goedart, J. J., Fleming, M., Travis, W. D., Williams, A. E., Rabkin, C. S., et al. (1996) SV40 and pleural mesotheliomas in humans. Cancer Epidemiol. Biomarkers Prev. 5, 473–475.

    CAS  PubMed  Google Scholar 

  12. Karlsen, F., Kalantari, M., Chitemerere, M., Johansson, B., and Hagmar, B. (1994) Modifications of human and viral deoxyribonucleic acid by formaldehyde fixation. Lab. Invest. 71, 604–611.

    CAS  PubMed  Google Scholar 

  13. Lednicky, J. A., Garcea, R. L., Bergsagel, D. J., and Butel, J. S. (1995) Natural simian virus 40 strains are present in human choroid plexus and ependymoma tumors. Virology 212, 710–717.

    Article  CAS  PubMed  Google Scholar 

  14. Pääbo, S. (1990) Amplifying ancient DNA, in PCR Protocols: A Guide to Methods and Applications (Innis, Gelfand, Sninsky, and White, eds.), Academic, San Diego, CA.

    Google Scholar 

  15. Agostini, H. T. and Stoner, G. L. (1995) Amplification of the complete polyomavirus JC genome from brain cerebrospinal fluid and urine using pre-PCR restriction enzyme digeston. J. Neurovirol. 1, 316–320.

    Article  CAS  PubMed  Google Scholar 

  16. Lednicky, J. A., Jafar, S., Wong, C., and Butel, J. S. (1997) High-fidelity PCR amplification of infectious copies of the complete simian virus 40 genome from plasmids and virus-infected cell lysates. Gene 184, 189–195.

    Article  CAS  PubMed  Google Scholar 

  17. Laghi, L., Randolph, A. E., Chauhan, D. P., Marra, G., Major, E. O., Neel, J. V., et al. (1999) JC virus DNA is present in the mucosa of the human colon and in colorectal cancers. Proc. Natl. Acad. Sci. USA 96, 7484–7489.

    Article  CAS  PubMed  Google Scholar 

  18. Lednicky, J. A. and Butel, J. S. (1997) Tissue culture adaptation of natural isolates of simian virus 40: changes occur in viral regulatory region but not in carboxy-terminal domain of large T-antigen. J. Gen. Virol. 78, 1697–1705.

    CAS  PubMed  Google Scholar 

  19. Lednicky, J. A., Arrington, A. S., Stewart, A. R., Dai, X. M., Wong, C., Jafar, S., et al. (1998) Natural isolates of simian virus 40 from immunocompromised monkeys display extensive genetic heterogeneity: new implications for polyomavirus disease. J. Virol. 72, 3980–3990.

    CAS  PubMed  Google Scholar 

  20. Stewart, A. R., Lednicky, J. A., Benzick, U. S., Tevethia, M. J., and Butel, J. S. (1996) Identification of a variable region at the carboxy terminus of SV40 large T-antigen. Virology 221, 355–361.

    Article  CAS  PubMed  Google Scholar 

  21. Stewart, A. R., Lednicky, J. A., and Butel, J. S. (1998) Sequence analyses of human tumor-associated SV40 DNAs and SV40 viral isolates from monkeys and humans. NeuroVirol. 4, 182–193.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2001 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Lednicky, J.A., Garcea, R.L. (2001). Detection of SV40 DNA Sequences in Human Tissue. In: Raptis, L. (eds) SV40 Protocols. Methods in Molecular Biology™, vol 165. Humana Press. https://doi.org/10.1385/1-59259-117-5:257

Download citation

  • DOI: https://doi.org/10.1385/1-59259-117-5:257

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-653-6

  • Online ISBN: 978-1-59259-117-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics