Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 May;33(5):1263–1266. doi: 10.1128/jcm.33.5.1263-1266.1995

Early detection of human cytomegalovirus viremia in bone marrow transplant recipients by DNA amplification.

F S Nolte 1, R K Emmens 1, C Thurmond 1, P S Mitchell 1, C Pascuzzi 1, S M Devine 1, R Saral 1, J R Wingard 1
PMCID: PMC228142  PMID: 7615738

Abstract

Surveillance blood cultures for human cytomegalovirus (HCMV) are commonly used to identify the bone marrow transplant (BMT) recipients with the highest risk of serious HCMV disease and for whom early interventional ganciclovir therapy would be beneficial. We monitored 36 allogeneic BMT recipients weekly for the presence of HCMV in the blood from 0 to 100 days posttransplantation. Viable HCMV in leukocytes (WBC) was detected by shell vial and tube culture methods. HCMV DNA in WBC and plasma was detected by PCR and DNA hybridization using primers and a probe from the EcoRI fragment D region of HCMV AD169. A uracil-N-glycosylase-dUTP PCR protocol was used to prevent false-positive results due to amplicon carryover. Seventeen patients had multiple consecutive positive samples containing HCMV DNA in plasma or WBC. In 14 of 17 patients, HCMV was also detected by blood culture. HCMV DNA was detected sporadically in six patients, none of whom had positive cultures. One patient had HCMV viremia detected by WBC culture only. The remaining 12 patients had no positive PCR assays or blood cultures. For the patients with positive blood cultures, PCR detection of HCMV DNA in plasma preceded detection of HCMV in culture by a mean of 8 days and detection in WBC preceded detection in culture by 6 days. HCMV disease (interstitial pneumonia) was documented for two patients with viremia (blood culture and PCR positive) and one patient without viremia (blood culture and PCR negative). The earlier recognition of high-risk patients provided by detection of HCMV DNA in plasma or WBC may improve the efficacy of early interventional antiviral therapy.

Full Text

The Full Text of this article is available as a PDF (173.2 KB).

Selected References

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

  1. Bitsch A., Kirchner H., Dupke R., Bein G. Cytomegalovirus transcripts in peripheral blood leukocytes of actively infected transplant patients detected by reverse transcription-polymerase chain reaction. J Infect Dis. 1993 Mar;167(3):740–743. doi: 10.1093/infdis/167.3.740. [DOI] [PubMed] [Google Scholar]
  2. Boeckh M., Bowden R. A., Goodrich J. M., Pettinger M., Meyers J. D. Cytomegalovirus antigen detection in peripheral blood leukocytes after allogeneic marrow transplantation. Blood. 1992 Sep 1;80(5):1358–1364. [PubMed] [Google Scholar]
  3. Brytting M., Xu W., Wahren B., Sundqvist V. A. Cytomegalovirus DNA detection in sera from patients with active cytomegalovirus infections. J Clin Microbiol. 1992 Aug;30(8):1937–1941. doi: 10.1128/jcm.30.8.1937-1941.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Casareale D., Pottathil R., Diaco R. Improved blood sample processing for PCR. PCR Methods Appl. 1992 Nov;2(2):149–153. doi: 10.1101/gr.2.2.149. [DOI] [PubMed] [Google Scholar]
  5. Einsele H., Ehninger G., Steidle M., Vallbracht A., Müller M., Schmidt H., Saal J. G., Waller H. D., Müller C. A. Polymerase chain reaction to evaluate antiviral therapy for cytomegalovirus disease. Lancet. 1991 Nov 9;338(8776):1170–1172. doi: 10.1016/0140-6736(91)92032-w. [DOI] [PubMed] [Google Scholar]
  6. Einsele H., Steidle M., Vallbracht A., Saal J. G., Ehninger G., Müller C. A. Early occurrence of human cytomegalovirus infection after bone marrow transplantation as demonstrated by the polymerase chain reaction technique. Blood. 1991 Mar 1;77(5):1104–1110. [PubMed] [Google Scholar]
  7. Gerdes J. C., Spees E. K., Fitting K., Hiraki J., Sheehan M., Duda D., Jarvi T., Roehl C., Robertson A. D. Prospective study utilizing a quantitative polymerase chain reaction for detection of cytomegalovirus DNA in the blood of renal transplant patients. Transplant Proc. 1993 Feb;25(1 Pt 2):1411–1413. [PubMed] [Google Scholar]
  8. Gerna G., Zipeto D., Parea M., Revello M. G., Silini E., Percivalle E., Zavattoni M., Grossi P., Milanesi G. Monitoring of human cytomegalovirus infections and ganciclovir treatment in heart transplant recipients by determination of viremia, antigenemia, and DNAemia. J Infect Dis. 1991 Sep;164(3):488–498. doi: 10.1093/infdis/164.3.488. [DOI] [PubMed] [Google Scholar]
  9. Gleaves C. A., Smith T. F., Shuster E. A., Pearson G. R. Comparison of standard tube and shell vial cell culture techniques for the detection of cytomegalovirus in clinical specimens. J Clin Microbiol. 1985 Feb;21(2):217–221. doi: 10.1128/jcm.21.2.217-221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goodrich J. M., Bowden R. A., Fisher L., Keller C., Schoch G., Meyers J. D. Ganciclovir prophylaxis to prevent cytomegalovirus disease after allogeneic marrow transplant. Ann Intern Med. 1993 Feb 1;118(3):173–178. doi: 10.7326/0003-4819-118-3-199302010-00003. [DOI] [PubMed] [Google Scholar]
  11. Howell C. L., Miller M. J., Martin W. J. Comparison of rates of virus isolation from leukocyte populations separated from blood by conventional and Ficoll-Paque/Macrodex methods. J Clin Microbiol. 1979 Oct;10(4):533–537. doi: 10.1128/jcm.10.4.533-537.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hsia K., Spector D. H., Lawrie J., Spector S. A. Enzymatic amplification of human cytomegalovirus sequences by polymerase chain reaction. J Clin Microbiol. 1989 Aug;27(8):1802–1809. doi: 10.1128/jcm.27.8.1802-1809.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ishigaki S., Takeda M., Kura T., Ban N., Saitoh T., Sakamaki S., Watanabe N., Kohgo Y., Niitsu Y. Cytomegalovirus DNA in the sera of patients with cytomegalovirus pneumonia. Br J Haematol. 1991 Oct;79(2):198–204. doi: 10.1111/j.1365-2141.1991.tb04522.x. [DOI] [PubMed] [Google Scholar]
  14. Jiwa N. M., Van Gemert G. W., Raap A. K., Van de Rijke F. M., Mulder A., Lens P. F., Salimans M. M., Zwaan F. E., Van Dorp W., Van der Ploeg M. Rapid detection of human cytomegalovirus DNA in peripheral blood leukocytes of viremic transplant recipients by the polymerase chain reaction. Transplantation. 1989 Jul;48(1):72–76. doi: 10.1097/00007890-198907000-00017. [DOI] [PubMed] [Google Scholar]
  15. Longo M. C., Berninger M. S., Hartley J. L. Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions. Gene. 1990 Sep 1;93(1):125–128. doi: 10.1016/0378-1119(90)90145-h. [DOI] [PubMed] [Google Scholar]
  16. Marsano L., Perrillo R. P., Flye M. W., Hanto D. W., Spitzer E. D., Thomas J. R., Murray P. R., Windus D. W., Brunt E. M., Storch G. A. Comparison of culture and serology for the diagnosis of cytomegalovirus infection in kidney and liver transplant recipients. J Infect Dis. 1990 Mar;161(3):454–461. doi: 10.1093/infdis/161.3.454. [DOI] [PubMed] [Google Scholar]
  17. Meyers J. D., Ljungman P., Fisher L. D. Cytomegalovirus excretion as a predictor of cytomegalovirus disease after marrow transplantation: importance of cytomegalovirus viremia. J Infect Dis. 1990 Aug;162(2):373–380. doi: 10.1093/infdis/162.2.373. [DOI] [PubMed] [Google Scholar]
  18. Meyers J. D., Reed E. C., Shepp D. H., Thornquist M., Dandliker P. S., Vicary C. A., Flournoy N., Kirk L. E., Kersey J. H., Thomas E. D. Acyclovir for prevention of cytomegalovirus infection and disease after allogeneic marrow transplantation. N Engl J Med. 1988 Jan 14;318(2):70–75. doi: 10.1056/NEJM198801143180202. [DOI] [PubMed] [Google Scholar]
  19. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  20. Schmidt G. M., Horak D. A., Niland J. C., Duncan S. R., Forman S. J., Zaia J. A. A randomized, controlled trial of prophylactic ganciclovir for cytomegalovirus pulmonary infection in recipients of allogeneic bone marrow transplants; The City of Hope-Stanford-Syntex CMV Study Group. N Engl J Med. 1991 Apr 11;324(15):1005–1011. doi: 10.1056/NEJM199104113241501. [DOI] [PubMed] [Google Scholar]
  21. Spector S. A., Merrill R., Wolf D., Dankner W. M. Detection of human cytomegalovirus in plasma of AIDS patients during acute visceral disease by DNA amplification. J Clin Microbiol. 1992 Sep;30(9):2359–2365. doi: 10.1128/jcm.30.9.2359-2365.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wingard J. R., Piantadosi S., Burns W. H., Zahurak M. L., Santos G. W., Saral R. Cytomegalovirus infections in bone marrow transplant recipients given intensive cytoreductive therapy. Rev Infect Dis. 1990 Sep-Oct;12 (Suppl 7):S793–S804. doi: 10.1093/clinids/12.supplement_7.s793. [DOI] [PubMed] [Google Scholar]
  23. Winston D. J., Ho W. G., Bartoni K., Du Mond C., Ebeling D. F., Buhles W. C., Champlin R. E. Ganciclovir prophylaxis of cytomegalovirus infection and disease in allogeneic bone marrow transplant recipients. Results of a placebo-controlled, double-blind trial. Ann Intern Med. 1993 Feb 1;118(3):179–184. doi: 10.7326/0003-4819-118-3-199302010-00004. [DOI] [PubMed] [Google Scholar]
  24. Winston D. J., Ho W. G., Champlin R. E. Cytomegalovirus infections after allogeneic bone marrow transplantation. Rev Infect Dis. 1990 Sep-Oct;12 (Suppl 7):S776–S792. doi: 10.1093/clinids/12.supplement_7.s776. [DOI] [PubMed] [Google Scholar]
  25. Wolf D. G., Spector S. A. Early diagnosis of human cytomegalovirus disease in transplant recipients by DNA amplification in plasma. Transplantation. 1993 Aug;56(2):330–334. doi: 10.1097/00007890-199308000-00014. [DOI] [PubMed] [Google Scholar]
  26. van der Bij W., Torensma R., van Son W. J., Anema J., Schirm J., Tegzess A. M., The T. H. Rapid immunodiagnosis of active cytomegalovirus infection by monoclonal antibody staining of blood leucocytes. J Med Virol. 1988 Jun;25(2):179–188. doi: 10.1002/jmv.1890250208. [DOI] [PubMed] [Google Scholar]

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

RESOURCES