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. 1996 Sep;34(9):2085–2088. doi: 10.1128/jcm.34.9.2085-2088.1996

Monitoring of cytomegalovirus infection and disease in bone marrow recipients by reverse transcription-PCR and comparison with PCR and blood and urine cultures.

J Gozlan 1, J P Laporte 1, S Lesage 1, M Labopin 1, A Najman 1, N C Gorin 1, J C Petit 1
PMCID: PMC229194  PMID: 8862562

Abstract

Preemptive therapy is a promising strategy for the prevention of serious cytomegalovirus (CMV) disease after bone marrow (BM) transplantation but requires relevant diagnostic tests. We compared the clinical value of a reverse transcription (RT)-PCR method, which detected a late viral mRNA in peripheral blood leukocytes (PBL), with a PCR method that detected the viral DNA in PBL and with viral culture from leukocytes and urine for the diagnosis of symptomatic CMV infection after BM transplantation. Forty-five consecutive BM recipients were prospectively tested at weekly intervals by the four methods. CMV infection, demonstrated either by the culture of CMV or by repeated detection of viral DNA, was observed in 28 patients, but only 14 developed CMV-related clinical symptoms. The clinical sensitivity and specificity of each technique for detection of symptomatic infection were, respectively, 36 and 74% for urine culture, 43 and 84% for leukocyte culture, 100 and 65% for PCR, and 71 and 94% for RT-PCR. Although PCR detection of DNA in PBL was the earliest and most sensitive technique for the diagnosis of CMV infection, RT-PCR was more predictive of the onset of CMV-related clinical symptoms. These data suggest that both molecular methods should be used for identifying BM recipients at highest risk of CMV disease.

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Selected References

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  1. Alpert G., Mazeron M. C., Colimon R., Plotkin S. Rapid detection of human cytomegalovirus in the urine of humans. J Infect Dis. 1985 Sep;152(3):631–633. doi: 10.1093/infdis/152.3.631. [DOI] [PubMed] [Google Scholar]
  2. Altman D. G., Bland J. M. Diagnostic tests 2: Predictive values. BMJ. 1994 Jul 9;309(6947):102–102. doi: 10.1136/bmj.309.6947.102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Altman D. G., Bland J. M. Diagnostic tests. 1: Sensitivity and specificity. BMJ. 1994 Jun 11;308(6943):1552–1552. doi: 10.1136/bmj.308.6943.1552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Apperley J. F., Dowding C., Hibbin J., Buiter J., Matutes E., Sissons P. J., Gordon M., Goldman J. M. The effect of cytomegalovirus on hemopoiesis: in vitro evidence for selective infection of marrow stromal cells. Exp Hematol. 1989 Jan;17(1):38–45. [PubMed] [Google Scholar]
  5. Bilgrami S., Almeida G. D., Quinn J. J., Tuck D., Bergstrom S., Dainiak N., Poliquin C., Ascensao J. L. Pancytopenia in allogeneic marrow transplant recipients: role of cytomegalovirus. Br J Haematol. 1994 Jun;87(2):357–362. doi: 10.1111/j.1365-2141.1994.tb04922.x. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Dankner W. M., McCutchan J. A., Richman D. D., Hirata K., Spector S. A. Localization of human cytomegalovirus in peripheral blood leukocytes by in situ hybridization. J Infect Dis. 1990 Jan;161(1):31–36. doi: 10.1093/infdis/161.1.31. [DOI] [PubMed] [Google Scholar]
  8. Einsele H., Ehninger G., Hebart H., Wittkowski K. M., Schuler U., Jahn G., Mackes P., Herter M., Klingebiel T., Löffler J. Polymerase chain reaction monitoring reduces the incidence of cytomegalovirus disease and the duration and side effects of antiviral therapy after bone marrow transplantation. Blood. 1995 Oct 1;86(7):2815–2820. [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Emanuel D., Cunningham I., Jules-Elysee K., Brochstein J. A., Kernan N. A., Laver J., Stover D., White D. A., Fels A., Polsky B. Cytomegalovirus pneumonia after bone marrow transplantation successfully treated with the combination of ganciclovir and high-dose intravenous immune globulin. Ann Intern Med. 1988 Nov 15;109(10):777–782. doi: 10.7326/0003-4819-109-10-777. [DOI] [PubMed] [Google Scholar]
  12. First L. R., Smith B. R., Lipton J., Nathan D. G., Parkman R., Rappeport J. M. Isolated thrombocytopenia after allogeneic bone marrow transplantation: existence of transient and chronic thrombocytopenic syndromes. Blood. 1985 Feb;65(2):368–374. [PubMed] [Google Scholar]
  13. Gerna G., Zipeto D., Percivalle E., Parea M., Revello M. G., Maccario R., Peri G., Milanesi G. Human cytomegalovirus infection of the major leukocyte subpopulations and evidence for initial viral replication in polymorphonuclear leukocytes from viremic patients. J Infect Dis. 1992 Dec;166(6):1236–1244. doi: 10.1093/infdis/166.6.1236. [DOI] [PubMed] [Google Scholar]
  14. Goodrich J. M., Mori M., Gleaves C. A., Du Mond C., Cays M., Ebeling D. F., Buhles W. C., DeArmond B., Meyers J. D. Early treatment with ganciclovir to prevent cytomegalovirus disease after allogeneic bone marrow transplantation. N Engl J Med. 1991 Dec 5;325(23):1601–1607. doi: 10.1056/NEJM199112053252303. [DOI] [PubMed] [Google Scholar]
  15. Gorin N. C., Lopez M., Laporte J. P., Quittet P., Lesage S., Lemoine F., Berenson R. J., Isnard F., Grande M., Stachowiak J. Preparation and successful engraftment of purified CD34+ bone marrow progenitor cells in patients with non-Hodgkin's lymphoma. Blood. 1995 Mar 15;85(6):1647–1654. [PubMed] [Google Scholar]
  16. Gozlan J., Caburet F., Tancrede C., Petit J. C. A reverse polymerase chain reaction method for detection of human cytomegalovirus late transcripts in cells infected in vitro. J Virol Methods. 1992 Oct;40(1):1–10. doi: 10.1016/0166-0934(92)90002-u. [DOI] [PubMed] [Google Scholar]
  17. Gozlan J., Salord J. M., Chouaïd C., Duvivier C., Picard O., Meyohas M. C., Petit J. C. Human cytomegalovirus (HCMV) late-mRNA detection in peripheral blood of AIDS patients: diagnostic value for HCMV disease compared with those of viral culture and HCMV DNA detection. J Clin Microbiol. 1993 Jul;31(7):1943–1945. doi: 10.1128/jcm.31.7.1943-1945.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
  19. Laporte J. P., Douay L., Lopez M., Labopin M., Jouet J. P., Lesage S., Stachowiak J., Fouillard L., Isnard F., Noel-Walter M. P. One hundred twenty-five adult patients with primary acute leukemia autografted with marrow purged by mafosfamide: a 10-year single institution experience. Blood. 1994 Dec 1;84(11):3810–3818. [PubMed] [Google Scholar]
  20. MacKintosh F. R., Adlish J., Hall S. W., St Jeor S., Smith E., Tavassoli M., Zanjani E. D. Suppression of normal human hematopoiesis by cytomegalovirus in vitro. Exp Hematol. 1993 Feb;21(2):243–250. [PubMed] [Google Scholar]
  21. Maciejewski J. P., Bruening E. E., Donahue R. E., Mocarski E. S., Young N. S., St Jeor S. C. Infection of hematopoietic progenitor cells by human cytomegalovirus. Blood. 1992 Jul 1;80(1):170–178. [PubMed] [Google Scholar]
  22. Meyer-König U., Serr A., von Laer D., Kirste G., Wolff C., Haller O., Neumann-Haefelin D., Hufert F. T. Human cytomegalovirus immediate early and late transcripts in peripheral blood leukocytes: diagnostic value in renal transplant recipients. J Infect Dis. 1995 Mar;171(3):705–709. doi: 10.1093/infdis/171.3.705. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Nolte F. S., Emmens R. K., Thurmond C., Mitchell P. S., Pascuzzi C., Devine S. M., Saral R., Wingard J. R. Early detection of human cytomegalovirus viremia in bone marrow transplant recipients by DNA amplification. J Clin Microbiol. 1995 May;33(5):1263–1266. doi: 10.1128/jcm.33.5.1263-1266.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Patel R., Smith T. F., Espy M., Portela D., Wiesner R. H., Krom R. A., Paya C. V. A prospective comparison of molecular diagnostic techniques for the early detection of cytomegalovirus in liver transplant recipients. J Infect Dis. 1995 Apr;171(4):1010–1014. doi: 10.1093/infdis/171.4.1010. [DOI] [PubMed] [Google Scholar]
  26. Roberts W. D., Weinberg K. I., Kohn D. B., Sender L., Parkman R., Lenarsky C. Granulocyte recovery in pediatric marrow transplant recipients treated with ganciclovir for cytomegalovirus infection. Am J Pediatr Hematol Oncol. 1993 Aug;15(3):320–323. [PubMed] [Google Scholar]
  27. 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]
  28. Saltzman R. L., Quirk M. R., Jordan M. C. Disseminated cytomegalovirus infection. Molecular analysis of virus and leukocyte interactions in viremia. J Clin Invest. 1988 Jan;81(1):75–81. doi: 10.1172/JCI113313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schmidt C. A., Oettle H., Wilborn F., Jessen J., Timm H., Schwerdtfeger R., Oertel J., Siegert W. Demonstration of cytomegalovirus after bone marrow transplantation by polymerase chain reaction, virus culture and antigen detection in buffy coat leukocytes. Bone Marrow Transplant. 1994 Jan;13(1):71–75. [PubMed] [Google Scholar]
  30. 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]
  31. Simmons P., Kaushansky K., Torok-Storb B. Mechanisms of cytomegalovirus-mediated myelosuppression: perturbation of stromal cell function versus direct infection of myeloid cells. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1386–1390. doi: 10.1073/pnas.87.4.1386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sing G. K., Ruscetti F. W. Preferential suppression of myelopoiesis in normal human bone marrow cells after in vitro challenge with human cytomegalovirus. Blood. 1990 May 15;75(10):1965–1973. [PubMed] [Google Scholar]
  33. Verdonck L. F., de Gast G. C., van Heugten H. G., Nieuwenhuis H. K., Dekker A. W. Cytomegalovirus infection causes delayed platelet recovery after bone marrow transplantation. Blood. 1991 Aug 1;78(3):844–848. [PubMed] [Google Scholar]
  34. Verdonck L. F., van Heugten H., de Gast G. C. Delay in platelet recovery after bone marrow transplantation: impact of cytomegalovirus infection. Blood. 1985 Oct;66(4):921–925. [PubMed] [Google Scholar]
  35. Wingard J. R., Chen D. Y., Burns W. H., Fuller D. J., Braine H. G., Yeager A. M., Kaiser H., Burke P. J., Graham M. L., Santos G. W. Cytomegalovirus infection after autologous bone marrow transplantation with comparison to infection after allogeneic bone marrow transplantation. Blood. 1988 May;71(5):1432–1437. [PubMed] [Google Scholar]
  36. 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]

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