Abstract
A method for rapid identification of human cytomegalovirus (HCMV) was developed with biotinylated DNA probes. BamHI restriction fragments from HCMV strain AD169 were selected and tested for their ability to detect virus in patient urine samples. All probes detected 30 pg of HCMV AD169 DNA. The BamHI B fragment detected 15 of 29 cell-culture-positive samples (sensitivity, 52%). There were four samples which were probe positive and cell culture negative (specificity, 87%). The D and H fragments used as combined probes detected 17 of 21 cell-culture-positive samples (sensitivity, 81%). There were five probe-positive and cell-culture-negative samples (specificity, 68.8%). The H fragment, when used alone, detected 11 of 14 culture-positive samples, and 5 samples were culture negative and probe positive. Sensitivity (78.6%) and specificity (76.2%) for the H fragment were similar to those for the combined probes, but the color intensity of the positive reactions detected by the H fragment alone was lower. There was unexpected cross-reactivity with herpes simplex type 1 and 2 controls when the combined D and H probes were used. Specific hybridization was demonstrated between subfragments of the HCMV BamHI D fragment and the herpes simplex virus type 1 EcoRI M fragment.
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- Baer R., Bankier A. T., Biggin M. D., Deininger P. L., Farrell P. J., Gibson T. J., Hatfull G., Hudson G. S., Satchwell S. C., Séguin C. DNA sequence and expression of the B95-8 Epstein-Barr virus genome. Nature. 1984 Jul 19;310(5974):207–211. doi: 10.1038/310207a0. [DOI] [PubMed] [Google Scholar]
- Bayer E. A., Wilchek M. The use of the avidin-biotin complex as a tool in molecular biology. Methods Biochem Anal. 1980;26:1–45. doi: 10.1002/9780470110461.ch1. [DOI] [PubMed] [Google Scholar]
- Beneke J. S., Tegtmeier G. E., Alter H. J., Luetkemeyer R. B., Solomon R., Bayer W. L. Relation of titers of antibodies to CMV in blood donors to the transmission of cytomegalovirus infection. J Infect Dis. 1984 Dec;150(6):883–888. doi: 10.1093/infdis/150.6.883. [DOI] [PubMed] [Google Scholar]
- Brigati D. J., Myerson D., Leary J. J., Spalholz B., Travis S. Z., Fong C. K., Hsiung G. D., Ward D. C. Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probes. Virology. 1983 Apr 15;126(1):32–50. doi: 10.1016/0042-6822(83)90460-9. [DOI] [PubMed] [Google Scholar]
- Chandler S. H., Alexander E. R., Holmes K. K. Epidemiology of cytomegaloviral infection in a heterogeneous population of pregnant women. J Infect Dis. 1985 Aug;152(2):249–256. doi: 10.1093/infdis/152.2.249. [DOI] [PubMed] [Google Scholar]
- 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]
- Coen D. M., Fleming H. E., Jr, Leslie L. K., Retondo M. J. Sensitivity of arabinosyladenine-resistant mutants of herpes simplex virus to other antiviral drugs and mapping of drug hypersensitivity mutations to the DNA polymerase locus. J Virol. 1985 Feb;53(2):477–488. doi: 10.1128/jvi.53.2.477-488.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Colimon R., Somogyi T., Bertrand C., Michelson S. Demonstration of the colinearity of human cytomegalovirus genomes and construction of restriction maps of unknown isolates using cloned subgenomic fragments. J Gen Virol. 1985 Oct;66(Pt 10):2183–2198. doi: 10.1099/0022-1317-66-10-2183. [DOI] [PubMed] [Google Scholar]
- Costa R. H., Draper K. G., Kelly T. J., Wagner E. K. An unusual spliced herpes simplex virus type 1 transcript with sequence homology to Epstein-Barr virus DNA. J Virol. 1985 May;54(2):317–328. doi: 10.1128/jvi.54.2.317-328.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Diosi P., Moldovan E., Tomescu N. Latent cytomegalovirus infection in blood donors. Br Med J. 1969 Dec 13;4(5684):660–662. doi: 10.1136/bmj.4.5684.660. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drew W. L., Sweet E. S., Miner R. C., Mocarski E. S. Multiple infections by cytomegalovirus in patients with acquired immunodeficiency syndrome: documentation by Southern blot hybridization. J Infect Dis. 1984 Dec;150(6):952–953. doi: 10.1093/infdis/150.6.952. [DOI] [PubMed] [Google Scholar]
- Fiala M., Payne J. E., Berne T. V., Moore T. C., Henle W., Montgomerie J. Z., Chatterjee S. N., Guze L. B. Epidemiology of cytomegalovirus infection after transplantation and immunosuppression. J Infect Dis. 1975 Oct;132(4):421–433. doi: 10.1093/infdis/132.4.421. [DOI] [PubMed] [Google Scholar]
- Fung J. C., Shanley J., Tilton R. C. Comparison of the detection of herpes simplex virus in direct clinical specimens with herpes simplex virus-specific DNA probes and monoclonal antibodies. J Clin Microbiol. 1985 Nov;22(5):748–753. doi: 10.1128/jcm.22.5.748-753.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibbs J. S., Chiou H. C., Hall J. D., Mount D. W., Retondo M. J., Weller S. K., Coen D. M. Sequence and mapping analyses of the herpes simplex virus DNA polymerase gene predict a C-terminal substrate binding domain. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7969–7973. doi: 10.1073/pnas.82.23.7969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibson T., Stockwell P., Ginsburg M., Barrell B. Homology between two EBV early genes and HSV ribonucleotide reductase and 38K genes. Nucleic Acids Res. 1984 Jun 25;12(12):5087–5099. doi: 10.1093/nar/12.12.5087. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldin A. L., Sandri-Goldin R. M., Levine M., Glorioso J. C. Cloning of herpes simplex virus type 1 sequences representing the whole genome. J Virol. 1981 Apr;38(1):50–58. doi: 10.1128/jvi.38.1.50-58.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Greenaway P. J., Oram J. D., Downing R. G., Patel K. Human cytomegalovirus DNA: BamHI, EcoRI and PstI restriction endonuclease cleavage maps. Gene. 1982 Jun;18(3):355–360. doi: 10.1016/0378-1119(82)90174-3. [DOI] [PubMed] [Google Scholar]
- Huang E. S., Pagano J. S. Human cytomegalovirus. II. Lack of relatedness to DNA of herpes simples I and II, Epstein-Barr virus, and nonhuman strains of cytomegalovirus. J Virol. 1974 Mar;13(3):642–645. doi: 10.1128/jvi.13.3.642-645.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Myerson D., Hackman R. C., Meyers J. D. Diagnosis of cytomegaloviral pneumonia by in situ hybridization. J Infect Dis. 1984 Aug;150(2):272–277. doi: 10.1093/infdis/150.2.272. [DOI] [PubMed] [Google Scholar]
- Pass R. F., Stagno S., Britt W. J., Alford C. A. Specific cell-mediated immunity and the natural history of congenital infection with cytomegalovirus. J Infect Dis. 1983 Dec;148(6):953–961. doi: 10.1093/infdis/148.6.953. [DOI] [PubMed] [Google Scholar]
- Pellett P. E., Biggin M. D., Barrell B., Roizman B. Epstein-Barr virus genome may encode a protein showing significant amino acid and predicted secondary structure homology with glycoprotein B of herpes simplex virus 1. J Virol. 1985 Dec;56(3):807–813. doi: 10.1128/jvi.56.3.807-813.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Preiksaitis J. K., Rosno S., Grumet C., Merigan T. C. Infections due to herpesviruses in cardiac transplant recipients: role of the donor heart and immunosuppressive therapy. J Infect Dis. 1983 Jun;147(6):974–981. doi: 10.1093/infdis/147.6.974. [DOI] [PubMed] [Google Scholar]
- Reynolds D. W., Stagno S., Hosty T. S., Tiller M., Alford C. A., Jr Maternal cytomegalovirus excretion and perinatal infection. N Engl J Med. 1973 Jul 5;289(1):1–5. doi: 10.1056/NEJM197307052890101. [DOI] [PubMed] [Google Scholar]
- Richardson W. P., Colvin R. B., Cheeseman S. H., Tolkoff-Rubin N. E., Herrin J. T., Cosimi A. B., Collins A. B., Hirsch M. S., McCluskey R. T., Russell P. S. Glomerulopathy associated with cytomegalovirus viremia in renal allografts. N Engl J Med. 1981 Jul 9;305(2):57–63. doi: 10.1056/NEJM198107093050201. [DOI] [PubMed] [Google Scholar]
- Richman D. D., Cleveland P. H., Redfield D. C., Oxman M. N., Wahl G. M. Rapid viral diagnosis. J Infect Dis. 1984 Mar;149(3):298–310. doi: 10.1093/infdis/149.3.298. [DOI] [PubMed] [Google Scholar]
- Rüger R., Bornkamm G. W., Fleckenstein B. Human cytomegalovirus DNA sequences with homologies to the cellular genome. J Gen Virol. 1984 Aug;65(Pt 8):1351–1364. doi: 10.1099/0022-1317-65-8-1351. [DOI] [PubMed] [Google Scholar]
- Sixbey J. W., Nedrud J. G., Raab-Traub N., Hanes R. A., Pagano J. S. Epstein-Barr virus replication in oropharyngeal epithelial cells. N Engl J Med. 1984 May 10;310(19):1225–1230. doi: 10.1056/NEJM198405103101905. [DOI] [PubMed] [Google Scholar]
- Spector D. H., Hock L., Tamashiro J. C. Cleavage maps for human cytomegalovirus DNA strain AD169 for restriction endonucleases EcoRI, BglII, and HindIII. J Virol. 1982 May;42(2):558–582. doi: 10.1128/jvi.42.2.558-582.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spector D. H., Vacquier J. P. Human cytomegalovirus (strain AD169) contains sequences related to the avian retrovirus oncogene v-myc. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3889–3893. doi: 10.1073/pnas.80.13.3889. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spector S. A., Hirata K. K., Newman T. R. Identification of multiple cytomegalovirus strains in homosexual men with acquired immunodeficiency syndrome. J Infect Dis. 1984 Dec;150(6):953–956. doi: 10.1093/infdis/150.6.953. [DOI] [PubMed] [Google Scholar]
- Spector S. A., Rua J. A., Spector D. H., McMillan R. Detection of human cytomegalovirus in clinical specimens by DNA-DNA hybridization. J Infect Dis. 1984 Jul;150(1):121–126. doi: 10.1093/infdis/150.1.121. [DOI] [PubMed] [Google Scholar]
- Stagno S., Reynolds D. W., Huang E. S., Thames S. D., Smith R. J., Alford C. A. Congenital cytomegalovirus infection. N Engl J Med. 1977 Jun 2;296(22):1254–1258. doi: 10.1056/NEJM197706022962203. [DOI] [PubMed] [Google Scholar]
- Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]




