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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
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.
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.
Denhardt, D.T. 1966. A membrane filter technique for the detection of complementary DNA. Biochem. Biophys. Res. Commun., 23, 641–646.
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.
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.
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.
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.
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.
Stalhandske, P. and Pettersson, U. 1982. Identification of DNA viruses by membrane filter hybridization. J. Clin. Microbiol., 15, 744–747.
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1984 ECSC, EEC, EAEC, Brussels-Luxembourg
About this chapter
Cite this chapter
Hyypia, T., Stalhandske, P., Vainionpaa, R., Halonen, P., Pettersson, U. (1984). Detection of Viral Nucleic Acids in Cell Cultures and in Clinical Specimens by Spot Hybridization. In: McNulty, M.S., McFerran, J.B. (eds) Recent Advances in Virus Diagnosis. Current Topics in Veterinary Medicine and Animal Science, vol 29. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-6039-8_8
Download citation
DOI: https://doi.org/10.1007/978-94-009-6039-8_8
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-009-6041-1
Online ISBN: 978-94-009-6039-8
eBook Packages: Springer Book Archive