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. 1993 Jul;31(7):1886–1891. doi: 10.1128/jcm.31.7.1886-1891.1993

Detection of adenovirus in clinical specimens by polymerase chain reaction and liquid-phase hybridization quantitated by time-resolved fluorometry.

J C Hierholzer 1, P E Halonen 1, P O Dahlen 1, P G Bingham 1, M M McDonough 1
PMCID: PMC265651  PMID: 8349768

Abstract

In addition to tests for the group-specific hexon antigen of adenoviruses, adenoviruses can be detected in clinical specimens by hybridization assays utilizing the widely shared base sequences of the region of the hexon gene that codes for the group-reactive determinants. We have developed a liquid-phase hybridization system with biotin- and europium-labeled probes which are reacted after DNA amplification of a 161-bp region of the hexon gene and which are quantitated by time-resolved (TR) fluorometry in streptavidin-coated microtiter wells. Polymerase chain reaction (PCR)-TR fluorometry is not a rapid test in the usual sense, but it is highly useful for specimens with inherent toxicity or with low virus yield, such as organ minces and specimens obtained late in the course of an illness. In a survey of 103 specimens tested by this method, including urine, stool, and tissue suspensions, the agreement with the hexon-specific TR fluoroimmunoassay antigen test for positive specimens was 100% and the sensitivity compared with that of virus culture was 91%. The PCR-TR fluorometry system was also shown to be advantageous as a quantitative measure of PCR products.

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Selected References

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  1. Allard A., Albinsson B., Wadell G. Detection of adenoviruses in stools from healthy persons and patients with diarrhea by two-step polymerase chain reaction. J Med Virol. 1992 Jun;37(2):149–157. doi: 10.1002/jmv.1890370214. [DOI] [PubMed] [Google Scholar]
  2. Allard A., Girones R., Juto P., Wadell G. Polymerase chain reaction for detection of adenoviruses in stool samples. J Clin Microbiol. 1990 Dec;28(12):2659–2667. doi: 10.1128/jcm.28.12.2659-2667.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dahlén P. O., Iitiä A. J., Skagius G., Frostell A., Nunn M. F., Kwiatkowski M. Detection of human immunodeficiency virus type 1 by using the polymerase chain reaction and a time-resolved fluorescence-based hybridization assay. J Clin Microbiol. 1991 Apr;29(4):798–804. doi: 10.1128/jcm.29.4.798-804.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dahlén P., Hurskainen P., Lövgren T., Hyypiä T. Time-resolved fluorometry for the identification of viral DNA in clinical specimens. J Clin Microbiol. 1988 Nov;26(11):2434–2436. doi: 10.1128/jcm.26.11.2434-2436.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dahlén P., Iitiä A., Mukkala V. M., Hurskainen P., Kwiatkowski M. The use of europium (Eu3+) labelled primers in PCR amplification of specific target DNA. Mol Cell Probes. 1991 Apr;5(2):143–149. doi: 10.1016/0890-8508(91)90009-9. [DOI] [PubMed] [Google Scholar]
  6. Diamandis E. P., Christopoulos T. K. Europium chelate labels in time-resolved fluorescence immunoassays and DNA hybridization assays. Anal Chem. 1990 Nov 15;62(22):1149A–1157A. doi: 10.1021/ac00221a001. [DOI] [PubMed] [Google Scholar]
  7. Gouvea V., Glass R. I., Woods P., Taniguchi K., Clark H. F., Forrester B., Fang Z. Y. Polymerase chain reaction amplification and typing of rotavirus nucleic acid from stool specimens. J Clin Microbiol. 1990 Feb;28(2):276–282. doi: 10.1128/jcm.28.2.276-282.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hemmilä I., Dakubu S., Mukkala V. M., Siitari H., Lövgren T. Europium as a label in time-resolved immunofluorometric assays. Anal Biochem. 1984 Mar;137(2):335–343. doi: 10.1016/0003-2697(84)90095-2. [DOI] [PubMed] [Google Scholar]
  9. Hierholzer J. C. Adenoviruses in the immunocompromised host. Clin Microbiol Rev. 1992 Jul;5(3):262–274. doi: 10.1128/cmr.5.3.262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hierholzer J. C., Johansson K. H., Anderson L. J., Tsou C. J., Halonen P. E. Comparison of monoclonal time-resolved fluoroimmunoassay with monoclonal capture-biotinylated detector enzyme immunoassay for adenovirus antigen detection. J Clin Microbiol. 1987 Sep;25(9):1662–1667. doi: 10.1128/jcm.25.9.1662-1667.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hurskainen P., Dahlén P., Ylikoski J., Kwiatkowski M., Siitari H., Lövgren T. Preparation of europium-labelled DNA probes and their properties. Nucleic Acids Res. 1991 Mar 11;19(5):1057–1061. doi: 10.1093/nar/19.5.1057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Knopf H. P., Papoian R. Preparation of europium-streptavidin in a time-resolved fluoroimmunoassay for interleukin-3. J Immunol Methods. 1991 Apr 25;138(2):233–236. doi: 10.1016/0022-1759(91)90171-b. [DOI] [PubMed] [Google Scholar]
  13. Norrby E. The structural and functional diversity of Adenovirus capsid components. J Gen Virol. 1969 Sep;5(2):221–236. doi: 10.1099/0022-1317-5-2-221. [DOI] [PubMed] [Google Scholar]
  14. Schröder J. P., Kuhlmann W. D. Detection of tetanus antitoxin using Eu(3+)-labeled anti-human immunoglobulin G monoclonal antibodies in a time-resolved fluorescence immunoassay. J Clin Microbiol. 1991 Jul;29(7):1504–1507. doi: 10.1128/jcm.29.7.1504-1507.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Scott-Taylor T. H., Hammond G. W. Conserved sequences of the adenovirus genome for detection of all human adenovirus types by hybridization. J Clin Microbiol. 1992 Jul;30(7):1703–1710. doi: 10.1128/jcm.30.7.1703-1710.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Suonpä M., Markela E., Ståhlberg T., Hemmilä I. Europium-labelled streptavidin as a highly sensitive universal label. Indirect time-resolved immunofluorometry of FSH and TSH. J Immunol Methods. 1992 May 18;149(2):247–253. doi: 10.1016/0022-1759(92)90256-s. [DOI] [PubMed] [Google Scholar]
  17. Syvänen A. C., Laaksonen M., Söderlund H. Fast quantification of nucleic acid hybrids by affinity-based hybrid collection. Nucleic Acids Res. 1986 Jun 25;14(12):5037–5048. doi: 10.1093/nar/14.12.5037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wolcott M. J. Advances in nucleic acid-based detection methods. Clin Microbiol Rev. 1992 Oct;5(4):370–386. doi: 10.1128/cmr.5.4.370. [DOI] [PMC free article] [PubMed] [Google Scholar]

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