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In Situ PCR

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Recombinant Protein Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 63))

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Abstract

Since we first reported our findings regarding the in situ amplification of HIV-1 gag gene in a HIV-1 infected cell line in 1990 (1), there has been an explosion of research in the area of in situ PCR. There are over 200 publications describing various forms of in situ gene amplifications (selected bibliography refs. 1–105), identifying various infectious agents (1–17,20,23,32,44,47–78,81–88,96), tumor marker genes (23,78–79,92), cytokines, growth factors and their receptors (37,39,40,65,77), and other genetic elements of interest, in peer reviewed journals (18,23,84,94). The polymerase chain reaction (PCR) method for amplification of defined gene sequences has proved to be a valuable tool not only for basic researchers but also for clinical scientists (2,4,5,20,23,79–82). Using even a minute amount of DNA or RNA and choosing a thermostable enzyme from a large variety of sources, one can amplify the amount of the gene of interest, which can be analyzed and/or sequenced. Thus, genes or portions of gene sequences present only in a small sample of cells or small fraction of mixed cellular populations can be examined. However, one of the major drawbacks of standard PCR technique is that the procedure does not allow the association of amplified signals of a specific gene segment with the histological cell type(s). For example, it would be advantageous to determine what types of cells in the peripheral blood circulation carry HIV-1 provirus at various stages of HIV-1 infection (2–6,62,78) and what percent of HIV-1-infected cells actually are expressing viral RNA (2–16,35,42). Similar approaches have been used to detect the presence of other gene sequences in tissue materials and pathological specimens (1–105).

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Authors and Affiliations

  1. Section of Molecular Retrovirology, Division of Infectious Diseases, Department of Medicine, The Dorrance H. Hamilton Laboratories, Thomas Jefferson University, Philadelphia, PA

    Omar Bagasra, Muhammad Mukhtar, Farida Shaheen & Roger J. Pomerantz

  2. Center of Human Retrovirology and Gene Therapy, Thomas Jefferson University, Philadelphia, PA

    Omar Bagasra, Muhammad Mukhtar, Farida Shaheen & Roger J. Pomerantz

Authors
  1. Omar Bagasra
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  2. Muhammad Mukhtar
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  3. Farida Shaheen
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  4. Roger J. Pomerantz
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Editor information

Editors and Affiliations

  1. Thomas Jefferson University, Philadelphia, PA

    Rocky S. Tuan

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© 1997 Humana Press Inc., Totowa, NJ

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Bagasra, O., Mukhtar, M., Shaheen, F., Pomerantz, R.J. (1997). In Situ PCR. In: Tuan, R.S. (eds) Recombinant Protein Protocols. Methods in Molecular Biology™, vol 63. Humana Press. https://doi.org/10.1385/0-89603-481-X:275

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  • DOI: https://doi.org/10.1385/0-89603-481-X:275

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-481-5

  • Online ISBN: 978-1-59259-549-5

  • eBook Packages: Springer Protocols

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