Advertisement

Detection of Viral Nucleic Acids in Cell Cultures and in Clinical Specimens by Spot Hybridization

  • T. Hyypia
  • P. Stalhandske
  • R. Vainionpaa
  • P. Halonen
  • U. Pettersson
Chapter
  • 60 Downloads
Part of the Current Topics in Veterinary Medicine and Animal Science book series (CTVM, volume 29)

Abstract

Nucleic acid hybridization was used for detection of adenovirus DNA directly in clinical specimens and enterovirus RNA in infected cells. DNA in 40 stool specimens, including 18 adenovirus positive and 22 adenovirus negative samples according to a highly sensitive radioimmunoassay (RIA), was spotted onto a nitrocellulose filter and hybridized with radioactively labelled adenovirus type 2 DNA probe. Sixteen specimens positive by RIA and one negative by RIA were positive in the hybridization test. A cloned partial copy of the coxsackie B3 virus genome was used to detect enteroviruses in infected cells. Cells infected with coxsackieviruses A and B, echo and polioviruses gave positive signals in the hybridization assay, whereas cells infected with other viruses did not.

Keywords

Stool Specimen Hybridization Assay Nitrocellulose Filter Viral Nucleic Acid Hybridization Test 
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. Brandsma, J. and Miller, G. 1980. Nucleic acid spot hybridization: rapid quantitative screening of lymphoid cell lines for Epstein-Barr viral DNA. Proc. Natl. Acad. Sci. USA, 77, 6851–6855.PubMedCrossRefGoogle Scholar
  2. Chou, S. and Merigan, T.C. 1983. Rapid ctetection and quantitation of human cytomegalovirus in urine through DNA hybridization. N. Engl. J. Med., 308, 921–925.PubMedCrossRefGoogle Scholar
  3. Denhardt, D.T. 1966. A membrane filter technique for the detection of complementary DNA. Biochem. Biophys. Res. Commun., 23, 641–646.PubMedCrossRefGoogle Scholar
  4. Flores, J., Boeggeman, E., Purcell, R.H., Sereno, M., Prez, I., White, L., Wyatt, R.G., Chanock, R.M. and Kapikian, A.Z. 1983. A dot hybridization assay for detection of rotavirus. Lancet, i, 555–559.CrossRefGoogle Scholar
  5. Halonen, P., Sarkkinen, H., Arstila, P., Hjertsson, E. and Torfason, E. 1980. Four-layer radioimmunoassay for detection of adenovirus in stool. J. Clin. Microbiol., 11, 614–617.PubMedGoogle Scholar
  6. Leary, J.J., Brigati, D.J. and Ward, D.C. 1983. Rapid and sensitive colorimetric method for visualising biotinlabelled DNA probes hybridized to DNA or RNA immobilised on nitrocellulose: Bio-blots. Proc. Natl. Acad. Sci. USA, 80, 4045–4049.PubMedCrossRefGoogle Scholar
  7. Owens, R.A. and Diener, T.O. 1981. Sensitive and rapid diagnosis of potato spindle tuber virioid disease by nucleic acid hybridization. Science 213, 670–672.PubMedCrossRefGoogle Scholar
  8. Rigby, P.W.J., Dieckmann, M., Rhodes, C. and Berg, P. 1977. Labelling deoxyribonucleic acid to high specific activity by nick translation with DNA-polymerase I. J. Mol. Biol., 113, 237–251.PubMedCrossRefGoogle Scholar
  9. Stalhandske, P. and Pettersson, U. 1982. Identification of DNA viruses by membrane filter hybridization. J. Clin. Microbiol., 15, 744–747.PubMedGoogle Scholar
  10. Virtanen, M., Palva, A., Laaksonen, M., Halonen, P., Soderlund, H. and Ranki, M. 1983. Novel test for rapid viral diagnosis: Detection of adenovirus in nasopharyngeal mucus aspirates by means of nucleic-acid sandwich hybridization. Lancet, i, 381–383.CrossRefGoogle Scholar

Copyright information

© ECSC, EEC, EAEC, Brussels-Luxembourg 1984

Authors and Affiliations

  • T. Hyypia
    • 1
  • P. Stalhandske
    • 2
  • R. Vainionpaa
    • 1
  • P. Halonen
    • 1
  • U. Pettersson
    • 2
  1. 1.Department of VirologyUniversity of TurkuTurkuFinland
  2. 2.Department of Medical GeneticsUniversity of UppsalaUppsalaSweden

Personalised recommendations