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Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1998 Dec;51(12):914–921. doi: 10.1136/jcp.51.12.914

Qualitative and semiquantitative polymerase chain reaction testing for cytomegalovirus DNA in serum allows prediction of CMV related disease in liver transplant recipients.

P C Evans 1, A Soin 1, T G Wreghitt 1, G J Alexander 1
PMCID: PMC501027  PMID: 10070333

Abstract

AIM: To identify cytomegalovirus (CMV) infection in liver transplant recipients by polymerase chain reaction (PCR) techniques and to separate the cases in which CMV related disease will occur, for whom treatment is indicated, from those in whom infection will remain innocuous. METHODS: The combination of qualitative and semiquantitative PCR of serum and urine was assessed to determine whether these assays can identify those at risk of CMV related disease and compared their performance with conventional approaches to diagnosis. RESULTS: Qualitative PCR of serum had superior specificity, sensitivity, and positive and negative predictive values compared with urine DEAFF (detection of early antigen fluorescent foci) and PCR of urine. All episodes of CMV related disease were associated with the presence of CMV DNA by PCR in serum or urine; CMV was detected before clinical onset in 70% and 60% of cases, respectively. The period over which CMV DNA could be detected was not correlated with CMV related disease. Both peak viral load and cumulative viral load estimated using a semiquantitative PCR method on serum samples positive by the qualitative method could be used to distinguish asymptomatic infection from CMV related disease with 100% specificity and sensitivity. In contrast semiquantitative PCR of urine was of little value. CONCLUSIONS: An approach based on PCR testing with a combination of qualitative and subsequently semiquantitative serum samples would improve the diagnosis of CMV infection and aid identification of those patients at risk of CMV related disease, allowing treatment to be targeted specifically.

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Selected References

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  1. Bowden R. A., Sayers M., Flournoy N., Newton B., Banaji M., Thomas E. D., Meyers J. D. Cytomegalovirus immune globulin and seronegative blood products to prevent primary cytomegalovirus infection after marrow transplantation. N Engl J Med. 1986 Apr 17;314(16):1006–1010. doi: 10.1056/NEJM198604173141602. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Converse P. J., Hess A. D., Tutschka P. J., Santos G. W. Effect of cyclosporine on the response of normal human lymphocytes to cytomegalovirus in vitro. Infect Immun. 1983 Sep;41(3):1226–1233. doi: 10.1128/iai.41.3.1226-1233.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Delgado R., Lumbreras C., Alba C., Pedraza M. A., Otero J. R., Gómez R., Moreno E., Noriega A. R., Payá C. V. Low predictive value of polymerase chain reaction for diagnosis of cytomegalovirus disease in liver transplant recipients. J Clin Microbiol. 1992 Jul;30(7):1876–1878. doi: 10.1128/jcm.30.7.1876-1878.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Demmler G. J., Buffone G. J., Schimbor C. M., May R. A. Detection of cytomegalovirus in urine from newborns by using polymerase chain reaction DNA amplification. J Infect Dis. 1988 Dec;158(6):1177–1184. doi: 10.1093/infdis/158.6.1177. [DOI] [PubMed] [Google Scholar]
  6. Drouet E., Michelson S., Denoyel G., Colimon R. Polymerase chain reaction detection of human cytomegalovirus in over 2000 blood specimens correlated with virus isolation and related to urinary virus excretion. J Virol Methods. 1993 Dec 31;45(3):259–276. doi: 10.1016/0166-0934(93)90112-5. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Evans P. C., Gray J., Wreghitt T. G., Alexander G. J. Optimisation of the polymerase chain reaction and dot-blot hybridisation for detecting cytomegalovirus DNA in urine: comparison with detection of early antigen fluorescent foci and culture. J Virol Methods. 1998 Jul;73(1):41–52. doi: 10.1016/s0166-0934(98)00039-1. [DOI] [PubMed] [Google Scholar]
  9. Fox A. S., Tolpin M. D., Baker A. L., Broelsch C. E., Whittington P. F., Jackson T., Thistlethwaite J. R., Stuart F. P. Seropositivity in liver transplant recipients as a predictor of cytomegalovirus disease. J Infect Dis. 1988 Feb;157(2):383–385. doi: 10.1093/infdis/157.2.383. [DOI] [PubMed] [Google Scholar]
  10. Gane E., Saliba F., Valdecasas G. J., O'Grady J., Pescovitz M. D., Lyman S., Robinson C. A. Randomised trial of efficacy and safety of oral ganciclovir in the prevention of cytomegalovirus disease in liver-transplant recipients. The Oral Ganciclovir International Transplantation Study Group [corrected]. Lancet. 1997 Dec 13;350(9093):1729–1733. doi: 10.1016/s0140-6736(97)05535-9. [DOI] [PubMed] [Google Scholar]
  11. Gerna G., Furione M., Baldanti F., Sarasini A. Comparative quantitation of human cytomegalovirus DNA in blood leukocytes and plasma of transplant and AIDS patients. J Clin Microbiol. 1994 Nov;32(11):2709–2717. doi: 10.1128/jcm.32.11.2709-2717.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  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. Karet F. E., Charnock-Jones D. S., Harrison-Woolrych M. L., O'Reilly G., Davenport A. P., Smith S. K. Quantification of mRNA in human tissue using fluorescent nested reverse-transcriptase polymerase chain reaction. Anal Biochem. 1994 Aug 1;220(2):384–390. doi: 10.1006/abio.1994.1354. [DOI] [PubMed] [Google Scholar]
  15. Kidd I. M., Fox J. C., Pillay D., Charman H., Griffiths P. D., Emery V. C. Provision of prognostic information in immunocompromised patients by routine application of the polymerase chain reaction for cytomegalovirus. Transplantation. 1993 Oct;56(4):867–871. doi: 10.1097/00007890-199310000-00018. [DOI] [PubMed] [Google Scholar]
  16. Kulski J. K. Quantitation of human cytomegalovirus DNA in leukocytes by end-point titration and duplex polymerase chain reaction. J Virol Methods. 1994 Sep;49(2):195–208. doi: 10.1016/0166-0934(94)90044-2. [DOI] [PubMed] [Google Scholar]
  17. Martin M., Mañez R., Linden P., Estores D., Torre-Cisneros J., Kusne S., Ondick L., Ptachcinski R., Irish W., Kisor D. A prospective randomized trial comparing sequential ganciclovir-high dose acyclovir to high dose acyclovir for prevention of cytomegalovirus disease in adult liver transplant recipients. Transplantation. 1994 Oct 15;58(7):779–785. doi: 10.1097/00007890-199410150-00005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miller M. J., Bovey S., Pado K., Bruckner D. A., Wagar E. A. Application of PCR to multiple specimen types for diagnosis of cytomegalovirus infection: comparison with cell culture and shell vial assay. J Clin Microbiol. 1994 Jan;32(1):5–10. doi: 10.1128/jcm.32.1.5-10.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mutimer D., Matyi-Toth A., Shaw J., Elias E., O'Donnell K., Stalhandske P. Patterns of viremia in liver transplant recipients with symptomatic cytomegalovirus infection. Transplantation. 1997 Jan 15;63(1):68–73. doi: 10.1097/00007890-199701150-00013. [DOI] [PubMed] [Google Scholar]
  20. Nyberg G., Bergström T., Blohmé I., Nordén G., Olofsson S., Ricksten A. Clinical evaluation in organ transplant patients of a polymerase chain reaction test for CMV DNA applied on white blood cells and serum. Transpl Int. 1994;7(6):428–433. doi: 10.1007/BF00346037. [DOI] [PubMed] [Google Scholar]
  21. Patel R., Smith T. F., Espy M., Wiesner R. H., Krom R. A., Portela D., Paya C. V. Detection of cytomegalovirus DNA in sera of liver transplant recipients. J Clin Microbiol. 1994 Jun;32(6):1431–1434. doi: 10.1128/jcm.32.6.1431-1434.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Prösch S., Kimel V., Dawydowa I., Krüger D. H. Monitoring of patients for cytomegalovirus after organ transplantation by centrifugation culture and PCR. J Med Virol. 1992 Dec;38(4):246–251. doi: 10.1002/jmv.1890380404. [DOI] [PubMed] [Google Scholar]
  23. Quinnan G. V., Jr, Kirmani N., Rook A. H., Manischewitz J. F., Jackson L., Moreschi G., Santos G. W., Saral R., Burns W. H. Cytotoxic t cells in cytomegalovirus infection: HLA-restricted T-lymphocyte and non-T-lymphocyte cytotoxic responses correlate with recovery from cytomegalovirus infection in bone-marrow-transplant recipients. N Engl J Med. 1982 Jul 1;307(1):7–13. doi: 10.1056/NEJM198207013070102. [DOI] [PubMed] [Google Scholar]
  24. Rawal B. K., Booth J. C., Fernando S., Butcher P. D., Powles R. L. Quantification of cytomegalovirus DNA in blood specimens from bone marrow transplant recipients by the polymerase chain reaction. J Virol Methods. 1994 Apr;47(1-2):189–202. doi: 10.1016/0166-0934(94)90077-9. [DOI] [PubMed] [Google Scholar]
  25. Reusser P., Riddell S. R., Meyers J. D., Greenberg P. D. Cytotoxic T-lymphocyte response to cytomegalovirus after human allogeneic bone marrow transplantation: pattern of recovery and correlation with cytomegalovirus infection and disease. Blood. 1991 Sep 1;78(5):1373–1380. [PubMed] [Google Scholar]
  26. Saliba F., Gugenheim J., Samuel D., Bismuth A., Mathieu D., Serres C., Bismuth H. Incidence of cytomegalovirus infection and effects of cytomegalovirus immune globulin prophylaxis after orthotopic liver transplantation. Transplant Proc. 1987 Oct;19(5):4081–4082. [PubMed] [Google Scholar]
  27. Schmidt C. A., Oettle H., Neuhaus P., Wiens M., Timm H., Wilborn F., Siegert W. Demonstration of cytomegalovirus by polymerase chain reaction after liver transplantation. Transplantation. 1993 Oct;56(4):872–874. doi: 10.1097/00007890-199310000-00019. [DOI] [PubMed] [Google Scholar]
  28. Schäfer P., Braun R. W., Möhring K., Henco K., Kang J., Wendland T., Kühn J. E. Quantitative determination of human cytomegalovirus target sequences in peripheral blood leukocytes by nested polymerase chain reaction and temperature gradient gel electrophoresis. J Gen Virol. 1993 Dec;74(Pt 12):2699–2707. doi: 10.1099/0022-1317-74-12-2699. [DOI] [PubMed] [Google Scholar]
  29. Singh N., Dummer J. S., Kusne S., Breinig M. K., Armstrong J. A., Makowka L., Starzl T. E., Ho M. Infections with cytomegalovirus and other herpesviruses in 121 liver transplant recipients: transmission by donated organ and the effect of OKT3 antibodies. J Infect Dis. 1988 Jul;158(1):124–131. doi: 10.1093/infdis/158.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Singh N., Yu V. L., Mieles L., Wagener M. M., Miner R. C., Gayowski T. High-dose acyclovir compared with short-course preemptive ganciclovir therapy to prevent cytomegalovirus disease in liver transplant recipients. A randomized trial. Ann Intern Med. 1994 Mar 1;120(5):375–381. doi: 10.7326/0003-4819-120-5-199403010-00004. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Stratta R. J., Shaefer M. S., Markin R. S., Wood R. P., Kennedy E. M., Langnas A. N., Reed E. C., Woods G. L., Donovan J. P., Pillen T. J. Clinical patterns of cytomegalovirus disease after liver transplantation. Arch Surg. 1989 Dec;124(12):1443–1450. doi: 10.1001/archsurg.1989.01410120093018. [DOI] [PubMed] [Google Scholar]
  33. Stratta R. J., Shaeffer M. S., Markin R. S., Wood R. P., Langnas A. N., Reed E. C., Donovan J. P., Woods G. L., Bradshaw K. A., Pillen T. J. Cytomegalovirus infection and disease after liver transplantation. An overview. Dig Dis Sci. 1992 May;37(5):673–688. doi: 10.1007/BF01296422. [DOI] [PubMed] [Google Scholar]
  34. Sutherland S., Bracken P., Wreghitt T. G., O'Grady J., Calne R. Y., Williams R. Donated organ as a source of cytomegalovirus in orthotopic liver transplantation. J Med Virol. 1992 Jul;37(3):170–173. doi: 10.1002/jmv.1890370304. [DOI] [PubMed] [Google Scholar]
  35. Winston D. J. Prevention of cytomegalovirus disease in transplant recipients. Lancet. 1995 Nov 25;346(8987):1380–1381. doi: 10.1016/s0140-6736(95)92401-9. [DOI] [PubMed] [Google Scholar]
  36. Winston D. J., Wirin D., Shaked A., Busuttil R. W. Randomised comparison of ganciclovir and high-dose acyclovir for long-term cytomegalovirus prophylaxis in liver-transplant recipients. Lancet. 1995 Jul 8;346(8967):69–74. doi: 10.1016/s0140-6736(95)92110-9. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Xu W., Sundqvist V. A., Brytting M., Linde A. Diagnosis of cytomegalovirus infections using polymerase chain reaction, virus isolation and serology. Scand J Infect Dis. 1993;25(3):311–316. doi: 10.3109/00365549309008504. [DOI] [PubMed] [Google Scholar]
  39. Yamaguchi Y., Hironaka T., Kajiwara M., Tateno E., Kita H., Hirai K. Increased sensitivity for detection of human cytomegalovirus in urine by removal of inhibitors for the polymerase chain reaction. J Virol Methods. 1992 May;37(2):209–218. doi: 10.1016/0166-0934(92)90048-i. [DOI] [PubMed] [Google Scholar]
  40. Zaia J. A., Forman S. J., Gallagher M. T., Vanderwal-Urbina E., Blume K. G. Prolonged human cytomegalovirus viremia following bone marrow transplantation. Transplantation. 1984 Mar;37(3):315–317. doi: 10.1097/00007890-198403000-00021. [DOI] [PubMed] [Google Scholar]
  41. Zipeto D., Revello M. G., Silini E., Parea M., Percivalle E., Zavattoni M., Milanesi G., Gerna G. Development and clinical significance of a diagnostic assay based on the polymerase chain reaction for detection of human cytomegalovirus DNA in blood samples from immunocompromised patients. J Clin Microbiol. 1992 Feb;30(2):527–530. doi: 10.1128/jcm.30.2.527-530.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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