Fast Multiplex Polymerase Chain Reaction on Boiled Clinical Samples for Rapid Diagnosis of Viral Infections

  • Christian Vandenvelde
  • Danièle Van Beers
Part of the Applied Virology Research book series (AVIR, volume 3)


In spite of the recent dramatic development of new, rapid methods for detection of viral infections, most laboratory diagnosis of viral disease still depends either on the growth of virus or on the detection of a specific serologic reaction to infection. As tissue culture is the most widely sensitive and flexible, it remains the golden standard. But, because no single monolayer type is sensitive to the growth of all viruses most laboratories choose an array of different tissue cultures and, depending on the suspected viruses, inoculate each specimen into those lines or strains most likely to give a positive result. For the most part each virus, or virus group, produces a characteristic cytopathic effect, and the skilled technologist can usually develop a tentative identification on the basis of cell tropism and morphology, giving the clinician useful information at the earliest possible moment. Final identification of virus isolates usually depends on serologic tests and sometimes on morphologic, biophysical, or biologic characterization. The polymerase chain reaction (PCR) has the potential to replace these laborious viral isolation methods, but problems such as specificity, reproducibility, rapidity, cost, and the need for a common sequence in all strains of a particular virus must be taken into account. The production of low and high molecular weight non-specific PCR products must be noted while using most published PCR protocols. The accumulation of such PCR-products depletes primers and deoxynucleotide-triphosphates from the reaction mixture and leads to competition for enzyme with the desired PCR-product. Diagnosis must then rely on final detection assays such as hybridization with radioactive oligonucleotide probes to guarantee sufficient specificity, sensitivity and reproducibility.


Polymerase Chain Reaction Protocol Polymerase Chain Reaction Procedure Long Control Region Cervical Scrape Polymerase Chain Reaction Yield 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Baker, C. C., Phelps, W. C, Lindgren, V,. Braun, M.J., Gonda, M.A. and Howley, P.M. (1987). J. Virol. 61, 962–971.PubMedGoogle Scholar
  2. Beckmann, A.M., Chu, J., Miller, B.A., Christiansen, A.E., McDougall, J.K. and Galloway, D.A. (1989). Proc. Eighth Intl. Papillomavirus Workshop. Taos, New Mexico.Google Scholar
  3. Breslauer, K.V., Frank, R., Blöcker, H. and Marky, L. A. (1986). Proc. Natl. Acad. Sci. USA. 83, 3746–3750.PubMedCrossRefGoogle Scholar
  4. Freier, M.S., Kierzek, R., Jaeger, J.A., Sugimoto, N., Caruthers, M. H., Neilson, T. and Turner, D. H. (1986). Proc. Natl. Acad. Sci. USA. 83, 9373–9377.PubMedCrossRefGoogle Scholar
  5. Goad, W. B. and Kanehisa, M. I. (1982). Nucleic Acids Res. 10, 265–278.PubMedCrossRefGoogle Scholar
  6. Hoch, H. and Chanutin, A. (1954). Arch. Biochem. Biophys. 51, 271–276.PubMedCrossRefGoogle Scholar
  7. Innis, M. A., Myambo, K. B., Gelfand, D. H. and Brow, M. A. D. (1988). Proc. Natl. Acad. Sci. USA. 85, 9436–9440.PubMedCrossRefGoogle Scholar
  8. Kaneko, S., Feinstone, S. M. and Miller, R. (1989). J. Clin. Microbiol. 27, 1930–1933.PubMedGoogle Scholar
  9. Kwok, S. and Higuchi, R. (1989). Nature 339, 237–238.PubMedCrossRefGoogle Scholar
  10. Larzul, D., Guigne, F., Sninsky, J., Mack, D., Bréchot, C. and Guesdon, J. (1988). J. Virol. Method. 20, 227–237.CrossRefGoogle Scholar
  11. Larzul, D., Chevrier, D., Thiers, V. and Guesdon, J. (1990). J. Virol Method. 27, 49–60.CrossRefGoogle Scholar
  12. Maniatis, T., Fritsch, E. F. and Sambrook, J. (1982). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor.Google Scholar
  13. Manos, M. M., Wright, D. K., Lewis, A. J., Wolinsky, S., Broker, T. R. and Ting, Y. (1989). Proc. Eighth Intl Papillomavirus Workshop,.Taos,New Mexico.Google Scholar
  14. Munoz, N., Bosch, X. and Keldor, J. M. (1988). Br. J. Cancer 57, 1–5.PubMedCrossRefGoogle Scholar
  15. Popescu, N. C., Di Paolo, J. A. and Amsbaugh, S. C. (1987). Cytogenet. Cell Genet. 44, 58–62.PubMedCrossRefGoogle Scholar
  16. Saiki, R. K., Bugawan, T. L., Horn, G. T., Mullis, K. B. and Erlich, H. A. (1986). Nature 324, 163–166.PubMedCrossRefGoogle Scholar
  17. Saiki, R. K. (1989). In PCR technology: principles and applications for DNA amplification. (H. A. Erlich, ed.). pp. 1–14. Stockton Press, London.Google Scholar
  18. Scotto, J., Hadchouel, M., Hery, C., Yvart, J., Tiollais, P. and Bréchot, C. (1983). Hepatology 3, 279–284.PubMedCrossRefGoogle Scholar
  19. Suggs, S. V., Hirose, T., Migaken, E. H., Kawashima, M. J., Johnson, K. I. and Wallace, R. B. (1981). In ICN-UCLA Symp Dev Biol Using Purified Genes. (D. D. Brown, ed.). pp. 683–693. Academic Press Inc, New-York.Google Scholar
  20. Tidy, J. A., Parry, G. C. N. and Ward, P. (1989). Lancet 1, 434.PubMedCrossRefGoogle Scholar
  21. Ulrich, P., Bath, R., Seto, B., Mack, D., Sninsky, J. and Vyas, G. (1989). J. Infect. Dis. 160, 37–43.PubMedCrossRefGoogle Scholar
  22. Vandenvelde, C., Scheen, R., Corazza, F. and Van Beers, D. (1991a). J. Exptl. Clin. Hematol. 33, 293–297.Google Scholar
  23. Vandenvelde, C., Scheen, R., Defoor, M., Duys, M., Dumon, J. and Van Beers, D. (1993). J. Virol. Method. 42, 251–264.CrossRefGoogle Scholar
  24. Vandenvelde, C., Scheen, R., Fondu, P. and Van Beers, D. (1991b). J. Exptl. Clin. Hematol. 33, 25–29.Google Scholar
  25. Vandenvelde, C., Scheen, R., Van Pachterbeke, C., Loriaux, C., Decelle, J., Hubert, T., Delhaye, C., Cattoor, J-P., Duys, M. and Van Beers, D. (1992). J. Med. Virol. 36, 279–282.PubMedCrossRefGoogle Scholar
  26. Vandenvelde, C. and Van Beers, D. (1992). J. Med. Virol. 38, 226–232.PubMedCrossRefGoogle Scholar
  27. Vandenvelde, C. and Van Beers, D. (1993). J. Med. Virol. 39, 273–277.PubMedCrossRefGoogle Scholar
  28. Vandenvelde, C., Verstraete, M. and Van Beers, D. (1990). J. Virol. Methods 30, 215–227.PubMedCrossRefGoogle Scholar
  29. Young, L. S., Bevan, J. S. and Johnson, M. A. (1989). Br. Med. J. 298, 14–18.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • Christian Vandenvelde
    • 1
  • Danièle Van Beers
    • 1
  1. 1.Hôpital Universitaire BrugmanUniversité Libre de BruxellesBruxellesBelgium

Personalised recommendations