Advertisement

Colorimetric In-Situ Hybridization in Clinical Virology: Development of Automated Technology

  • E. R. Unger
  • D. J. Brigati
Part of the Current Topics in Microbiology and Immunology book series (CT MICROBIOLOGY, volume 143)

Abstract

The goals of clinical virology, to generate accurate, sensitive and specific diagnoses as rapidly as possible, have not changed, but the methods to achieve these goals have undergone a revolution. The same techniques of molecular biology which proved so valuable in elucidating basic biologic properties of the viruses have been found to be applicable in a diagnostic setting. Through the adoption of monoclonal antibody and nucleic acid hybridization techniques, the clinical virology lab has been the indicator of things to come in microbiology in general. Because the culture of viruses is labor-intensive, expensive and often slow, the laboratory has been eager to adopt alternative methods to complement and streamline the “gold standard” of viral culture. The growing recognition of the clinical significance of noncultivatable viruses such as rota virus and human papilloma virus has also given impetus to the application of monoclonal antibodies and nucleic acid hybridization to these problems.

Keywords

Human Papilloma Virus Rota Virus Hybridization Assay Reagent Isolator Nucleic Acid Hybridization 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ambinder RF, Charache P, Staal S, Wright P, Forman M, Hayward SD, Hayward GS (1986) The vector homology problem in diagnostic nucleic acid hybridization of clinical samples. J Clin Microbiol 24: 16–20PubMedGoogle Scholar
  2. Brigati DJ, Myerson D, Leary JJ, Spalholz B, Travis SZ, Fong CKY, Hsiung GD, Ward DC (1983) Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probes. Virology 126: 32–50PubMedCrossRefGoogle Scholar
  3. Brigati DJ, Budgeon LR, Unger ER, Koebler D, Cuomo C, Kennedy T, Perdomo JMI (1988) Immunocytochemistry is automated. The development of a robotic workstation based upon the capillary action principle. J Histotechnol 11: 165–183Google Scholar
  4. Burns J, Redfern DRM, Esiri MM, McGee JOD (1986) Human and viral gene detection in routine paraffin embedded tissue by in situ hybridisation with biotinylated probes: viral localisation in herpes encephalitis. J Clin Pathol 39: 1066–1073PubMedCrossRefGoogle Scholar
  5. Grody WW, Cheng L, Lewin KJ (1987) In-situ viral DNA hybridization in diagnostic surgical pathology. Hum Pathol 18: 535–543PubMedCrossRefGoogle Scholar
  6. Highfield PE, Dougan G (1965) DNA probes for microbial diagnosis. Med Lab Sci 42: 352–360Google Scholar
  7. Landry ML, Fong CKY (1985) Nucleic acid hybridization in the diagnosis of viral infections. Clin Lab Med 5: 513–529PubMedGoogle Scholar
  8. Langer PR, Waldrop AA, Ward DC (1981) Enzymatic synthesis of biotin-labelled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci USA 18: 6633–6637Google Scholar
  9. Leary JJ, Brigati DJ, Ward DC (1983) Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci USA 80: 4045–4049Google Scholar
  10. Lewin R (1983) Genetic probes become even sharper. Science 229: 1167CrossRefGoogle Scholar
  11. Lewis FA, Griffiths S, Dunnicliff R, Wells M, Dudding N, Bird CC (1987) Sensitive in situ hybridisation technique using biotin-streptavidin-polyalkaline phosphatase complex. J Clin Pathol 40: 163–166PubMedCrossRefGoogle Scholar
  12. Palva A (1986) Microbial diagnostics by nucleic acid hybridization. Ann Clin Res 18: 327–336PubMedGoogle Scholar
  13. Singer RH, Lawrence JB, Villnave C (1986) Optimization of in situ hybridization using isotopie and non-isotopic detection methods. Biotechniques 4: 230–250Google Scholar
  14. Sklar J (1985) DNA hybridization in diagnostic pathology. Hum Pathol 16: 654–658PubMedCrossRefGoogle Scholar
  15. Unger ER, Budgeon LR, Myerson D, Brigati DJ (1986) Viral diagnosis by in-situ hybridization: description of a rapid simplified colorimetrie method. Am J Surg Pathol 10: 1–8PubMedCrossRefGoogle Scholar
  16. Unger ER, Leary J J, Ward DC, Brigati DJ (1987) Application of nucleic acid hybridization in clinical virology. In: Microbial antigenodiagnosis, vol 1. CRC Press, Boca RatonGoogle Scholar

Copyright information

© Springer-Verlag Berlin · Heidelberg 1989

Authors and Affiliations

  • E. R. Unger
    • 1
  • D. J. Brigati
    • 2
  1. 1.Department of Pathology and Laboratory MedicineEmory UniversityAtlantaUSA
  2. 2.Department of Pathology, The Milton S. Hershey Medical CenterThe Pennsylvania State UniversityHersheyUSA

Personalised recommendations