Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1985 May;21(5):730–733. doi: 10.1128/jcm.21.5.730-733.1985

Detection of adenovirus in nasopharyngeal specimens by radioactive and nonradioactive DNA probes.

T Hyypiä
PMCID: PMC271768  PMID: 3998103

Abstract

The presence of adenovirus DNA in clinical specimens was analyzed by nucleic acid hybridization assays by both radioactive and enzymatic detection systems. The sensitivity of the hybridization tests was in the range of 10 to 100 pg of homologous adenovirus DNA. Minimal background was noticed with unrelated viral and nonviral DNA. Twenty-four nasopharyngeal mucus aspirate specimens, collected from children with acute respiratory infection, were assayed in the hybridization tests and also by an enzyme immunoassay for adenovirus hexon antigen which was used as a reference test. Sixteen specimens positive by the enzyme immunoassay also were positive in the two nucleic acid hybridization tests, and the remaining eight specimens were negative in all of the tests. The results indicate that nucleid acid hybridization tests with both radioactive and nonradioactive probes can be used for diagnosis of microbial infections.

Full text

PDF
730

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berninger M., Hammer M., Hoyer B., Gerin J. L. An assay for the detection of the DNA genome of hepatitis B virus in serum. J Med Virol. 1982;9(1):57–68. doi: 10.1002/jmv.1890090109. [DOI] [PubMed] [Google Scholar]
  2. Brandsma J., Miller G. Nucleic acid spot hybridization: rapid quantitative screening of lymphoid cell lines for Epstein-Barr viral DNA. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6851–6855. doi: 10.1073/pnas.77.11.6851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chou S., Merigan T. C. Rapid detection and quantitation of human cytomegalovirus in urine through DNA hybridization. N Engl J Med. 1983 Apr 21;308(16):921–925. doi: 10.1056/NEJM198304213081603. [DOI] [PubMed] [Google Scholar]
  4. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  5. Echeverria P., Seriwatana J., Leksomboon U., Tirapat C., Chaicumpa W., Rowe B. Identification by DNA hybridisation of enterotoxigenic Escherichia coli in homes of children with diarrhoea. Lancet. 1984 Jan 14;1(8368):63–66. doi: 10.1016/s0140-6736(84)90001-1. [DOI] [PubMed] [Google Scholar]
  6. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 1971 Sep;8(9):871–874. doi: 10.1016/0019-2791(71)90454-x. [DOI] [PubMed] [Google Scholar]
  7. Flores J., Boeggeman E., Purcell R. H., Sereno M., Perez I., White L., Wyatt R. G., Chanock R. M., Kapikian A. Z. A dot hybridisation assay for detection of rotavirus. Lancet. 1983 Mar 12;1(8324):555–558. doi: 10.1016/s0140-6736(83)92811-8. [DOI] [PubMed] [Google Scholar]
  8. Franzén L., Westin G., Shabo R., Aslund L., Perlmann H., Persson T., Wigzell H., Pettersson U. Analysis of clinical specimens by hybridisation with probe containing repetitive DNA from Plasmodium falciparum. A novel approach to malaria diagnosis. Lancet. 1984 Mar 10;1(8376):525–528. doi: 10.1016/s0140-6736(84)90929-2. [DOI] [PubMed] [Google Scholar]
  9. Gilden D. H., Vafai A., Shtram Y., Becker Y., Devlin M., Wellish M. Varicella-zoster virus DNA in human sensory ganglia. Nature. 1983 Dec 1;306(5942):478–480. doi: 10.1038/306478a0. [DOI] [PubMed] [Google Scholar]
  10. Hyypiä T., Stålhandske P., Vainionpä R., Pettersson U. Detection of enteroviruses by spot hybridization. J Clin Microbiol. 1984 Mar;19(3):436–438. doi: 10.1128/jcm.19.3.436-438.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jalkanen M., Jalkanen S. Immunological detection of proteins after isoelectric focusing in thin layer agarose gel: a specific application for the characterization of immunoglobulin diversity. J Clin Lab Immunol. 1983 Apr;10(4):225–228. [PubMed] [Google Scholar]
  12. Langer P. R., Waldrop A. A., Ward D. C. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633–6637. doi: 10.1073/pnas.78.11.6633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leary J. J., Brigati D. J., Ward D. C. 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 U S A. 1983 Jul;80(13):4045–4049. doi: 10.1073/pnas.80.13.4045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pettersson U., Sambrook J. Amount of viral DNA in the genome of cells transformed by adenovirus type 2. J Mol Biol. 1973 Jan;73(1):125–130. doi: 10.1016/0022-2836(73)90164-2. [DOI] [PubMed] [Google Scholar]
  15. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  16. Sarkkinen H. K., Halonen P. E., Arstila P. P., Salmi A. A. Detection of respiratory syncytial, parainfluenza type 2, and adenovirus antigens by radioimmunoassay and enzyme immunoassay on nasopharyngeal specimens from children with acute respiratory disease. J Clin Microbiol. 1981 Feb;13(2):258–265. doi: 10.1128/jcm.13.2.258-265.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  18. Stålhandske P., Pettersson U. Identification of DNA viruses by membrane filter hybridization. J Clin Microbiol. 1982 Apr;15(4):744–747. doi: 10.1128/jcm.15.4.744-747.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Totten P. A., Holmes K. K., Handsfield H. H., Knapp J. S., Perine P. L., Falkow S. DNA hybridization technique for the detection of Neisseria gonorrhoeae in men with urethritis. J Infect Dis. 1983 Sep;148(3):462–471. doi: 10.1093/infdis/148.3.462. [DOI] [PubMed] [Google Scholar]
  20. Virtanen M., Palva A., Laaksonen M., Halonen P., Söderlund H., Ranki M. Novel test for rapid viral diagnosis: detection of adenovirus in nasopharyngeal mucus aspirates by means of nucleic-acid sandwich hybridisation. Lancet. 1983 Feb 19;1(8321):381–383. doi: 10.1016/s0140-6736(83)91500-3. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES