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Methods for the Detection of Cytomegalovirus in Glioblastoma Cells and Tissues

  • Charles S. CobbsEmail author
  • Lisa Matlaf
  • Lualhati E. Harkins
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1119)

Abstract

An increased awareness of the potential oncomodulatory properties of human cytomegalovirus (HCMV) has evolved over the last decade. We first reported the presence of HCMV in human glioblastomas, and subsequently these findings have been corroborated by other groups. However, some controversy has been associated with the immunohistochemical and in situ hybridization techniques used, since standard immunohistochemical and in situ hybridization techniques have been insufficient to detect low level HCMV antigens and nucleic acids in some tumor tissues. Here, we present detailed methods that can be used for the sensitive detection of low level HCMV antigens and nucleic acids in human glioblastoma specimens. Using these techniques, HCMV is frequently detected in frozen and formalin fixed paraffin-embedded tissue specimens. Furthermore, we demonstrate how human primary glioblastoma cells can be cultured in vitro, and how these cells can be used for detection of HCMV by immunofluorescence, in situ hybridization, western blot, and RT-PCR.

Key words

Human cytomegalovirus Glioblastoma Immunostaining PCR 

References

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Charles S. Cobbs
    • 1
    Email author
  • Lisa Matlaf
    • 2
  • Lualhati E. Harkins
    • 3
  1. 1.California Pacific Medical Center Research InstituteSan FranciscoUSA
  2. 2.California Pacific Medical Center Research InstituteSan FranciscoUSA
  3. 3.Department of Pathology and Laboratory MedicineBirmingham Veterans HospitalBirminghamUSA

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