Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Technical Report
  • Published:

A rapid monitoring system of human herpesviruses reactivation by LightCycler in stem cell transplantation

Bone Marrow Transplantation volume 28, pages 975–980 (2001)Cite this article

Abstract

To establish a practical monitoring system of human herpesviruses reactivation in patients undergoing stem cell transplantation, we developed a new, very rapid, highly sensitive, and quantitative PCR assay for accurate measurement of human cytomegalovirus (CMV), human herpesvirus 6 (HHV-6) and Epstein–Barr virus (EBV) DNA using LightCycler. The LightCycler system revealed that there was a linear correlation in the wide range of viral template DNA at the indicated number of PCR cycles. Peripheral blood cells were collected from 16 patients undergoing stem cell transplantation. The cut-off level of CMV and HHV-6 was assessed as 102 copies/μg and that of EBV as 103. High numbers of CMV genomes were detected in 3/13 patients after transplant, and reactivation of HHV-6 was frequently seen, whereas none of the patient showed an elevation of EBV genome copies until the end of the observation period. In the present study, the reactivation of β herpesviruses is associated with the occurrence of thrombotic microangiopathy (TMA) in two patients undergoing allogeneic BMT. Therefore, it may contribute in clarifying the pathological potential of human herpesviruses using a large number of clinical samples. Our results suggest that this system may be useful for monitoring viral reactivation. Bone Marrow Transplantation (2001) 28, 975–980.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. Ljungman P . Immune reconstitution and viral infections after stem cell transplantation Bone Marrow Transplant 1998 21 (Suppl. 2): S72–74

    Google Scholar 

  2. Wang F-Z, Dahl H, Linde A et al. Lymphotropic herpesvirus in allogeneic bone marrow transplantation Blood 1996 88: 3615–3620

    CAS  PubMed  Google Scholar 

  3. Broers AE, van Der Holt R, van Esser JW et al. Increased transplant-related morbidity and mortality in CMV-seropositive patients despite highly effective prevention of CMV disease after allogeneic T-cell-depleted stem cell transplantation Blood 2000 95: 2240–2245

    CAS  PubMed  Google Scholar 

  4. Wang F-Z, Linde A, Dahl H, Ljungman P . Human herpesvirus 6 infection inhibits specific lymphocyte proliferation and is related to lymphocytopenia after allogeneic bone marrow transplantation Bone Marrow Transplant 1999 24: 1201–1206

    Article  CAS  PubMed  Google Scholar 

  5. Bethge W, Beck R, Jahn G et al. Successful treatment of human herpesvirus-6 encephalitis after bone marrow transplantation Bone Marrow Transplant 1999 24: 1245–1248

    Article  CAS  PubMed  Google Scholar 

  6. Cone RW, Hackman R, Huang M-LW et al. Human herpesvirus 6 in lung tissue from patients with pneumonitis after bone marrow transplantation New Engl J Med 1993 329: 156–161

    Article  CAS  PubMed  Google Scholar 

  7. Cone RW, Huang ML, Grey L et al. Human herpesvirus 6 infections after bone marrow transplantation: clinical and virologic manifestations J Infect Dis 1999 179: 311–318

    Article  CAS  PubMed  Google Scholar 

  8. Lei KI, Chan LY, Chan WY et al. Quantitative analysis of circulating cell-free Epstein–Barr virus (EBV) DNA levels in patients with EBV-associated lymphoid malignancies Br J Haematol 2000 111: 239–246

    Article  CAS  PubMed  Google Scholar 

  9. Ohyashiki JH, Suzuki A, Aritaki K et al. Use of real-time PCR to monitor human herpesvirus 6 reactivation after allogeneic bone marrow transplantation Int J Mol Med 2000 6: 427–432

    CAS  PubMed  Google Scholar 

  10. Ohyashiki JH, Abe K, Ojima T et al. Quantification of human herpesvirus 6 in healthy volunteers and patients with lymphoproliferative disorders by PCR-ELISA Leukemia Res 1999 23: 625–630

    Article  CAS  Google Scholar 

  11. Ohyashiki JH, Yamamoto K, Suzuki A, Ohyashiki K . use of real-time PCR to monitor human β herpesviruses infection in hematologic disorders Jpn J Clin Hematol 2001 42: 468–471

    CAS  Google Scholar 

  12. Nagata H, Konno A, Kimura N et al. Characterization of novel natural killer (NK)-cell and gammadelta T-cell lines established from primary lesions of nasal T/NK-cell lymphomas associated with the Epstein–Barr virus Blood 2001 97: 708–713

    Article  CAS  PubMed  Google Scholar 

  13. Maeda Y, Teshima T, Yamada M et al. Monitoring of human herpesvirus after allogeneic peripheral blood stem cell transplantation and bone marrow transplantation Br J Haematol 1999 105: 295–302

    Article  CAS  PubMed  Google Scholar 

  14. Tanaka N, Kimura H, Hoshino Y et al. Monitoring for herpesviruses in unrelated cord blood transplantation Bone Marrow Transplant 2000 26: 1193–1197

    Article  CAS  PubMed  Google Scholar 

  15. Takemoto Y, Takatsuka H, Wada H et al. Evaluation of CMV/human herpes virus-6 positivity in bronchoalveolar lavage fluids as early detection of acute GVHD following BMT: evidence of a significant relationship Bone Marrow Transplant 2000 26: 77–81

    Article  CAS  PubMed  Google Scholar 

  16. Vogelsang GB . How I treat chronic graft-versus-host disease Blood 2001 97: 1196–1201

    Article  CAS  PubMed  Google Scholar 

  17. Zeigler Z, Kelton J, Moore J et al. Calpain activity in bone marrow transplant-associated thrombotic thrombocytopenic purpura Bone Marrow Transplant 1999 24: 641–645

    Article  CAS  PubMed  Google Scholar 

  18. Matsuda Y, Hara J, Miyoshi H et al. Thrombotic microangiopathy associated with reactivation of human herpesvirus 6 following high-dose chemotherapy with autologous bone marrow transplantation in young children Bone Marrow Transplant 1999 24: 919–923

    Article  CAS  PubMed  Google Scholar 

  19. Visseren FL, Verkerk MS, Bouter KP et al. Interleukin-6 production by endothelial cells after infection with influenza virus and cytomegalovirus J Lab Clin Med 1999 134: 623–630

    Article  CAS  PubMed  Google Scholar 

  20. Wu CA, Shanley JD . Chronic infection of human umbilical vein endothelial cells by human herpesvirus-6 J Gen Virol 1998 79: 1247–1256

    Article  CAS  PubMed  Google Scholar 

  21. Waldman WJ, Knight DA, Adams PW . Cytolytic activity against allogeneic human endothelia: resistance of cytomegalovirus-infected cells and virally activated lysis of uninfected cells Transplantation 1998 66: 67–77

    Article  CAS  PubMed  Google Scholar 

  22. Fish KN, Soderberg-Naucler C, Mills LK et al. Human cytomegalovirus persistently infects aortic endothelial cells J Virol 1998 72: 5661–5668

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Kahn M, Siegel-Acel D, Stenglein S et al. Efficient lytic infection of human arterial endothelial cells by human cytomegalovirus strains J Virol 2000 74: 7628–7635

    Article  Google Scholar 

  24. Ljungman P, Wang FZ, Clark DA et al. High levels of human herpesvirus 6 DNA in peripheral leukocytes are correlated to platelet engraftment and disease in allogeneic stem cell transplantation Br J Haematol 2000 111: 774–781

    CAS  PubMed  Google Scholar 

  25. Isomura H, Yoshida M, Namba H et al. Suppressive effects on human herpesvirus-6 on thrombopoietin-inducible megakaryocyte colony formation in vitro J Gen Virol 2000 81: 3663–3667

    Article  Google Scholar 

  26. Imbert-Marcille BM, Tang XW, Lepelletier D et al. Human herpesvirus 6 infection after autologous or allogeneic stem cell transplantation: a single-center prospective longitudinal study of 92 patients Clin Infect Dis 2000 31: 881–886

    Article  CAS  PubMed  Google Scholar 

  27. Nitsche A, Muller CW, Radonic A et al. Human herpesvirus 6A DNA is detected frequently in plasma but rarely in peripheral blood leukocytes of patients after bone marrow transplantation J Infect Dis 2001 183: 130–133

    Article  CAS  PubMed  Google Scholar 

  28. Ihira M, Yoshikawa T, Suzuki K et al. Correlation between human herpesvirus 6 and 7 infections after living related liver transplantation Microbiol Immunol 2001 45: 225–232

    Article  CAS  PubMed  Google Scholar 

  29. Lo YM, Chan LY, Chan AT et al. Quantitative and temporal correlation between circulating cell-free Epstein–Barr virus DNA and tumor recurrence in nasopharyngeal carcinoma Cancer Res 1999 59: 5452–5455

    CAS  PubMed  Google Scholar 

  30. Orii T, Ohkohchi N, Kikuchi H et al. Usefulness of quantitative real-time polymerase chain reaction in following up patients with Epstein-Barr virus infection after liver transplantation Clin Transplant 2000 4: 308–317

    Article  Google Scholar 

  31. Zhou CF, Abe K, Wang P et al. Efficient propagation of human herpesvirus 6B in a T cell line, derived from a patient with adult T cell leukemia/lymphoma Microbiol Immunol 1999 43: 425–436

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are indebted to Prof J Patrick Barron of the International Medical Communications Center of Tokyo Medical University for his review of this manuscript. The authors wish to thank Dr Naoko Shoji, Ms Ayako Hirota and Ms Michiko Kimura for their help.

Author information

Authors and Affiliations

  1. Department of Pediatrics, Tokyo Medical University, Tokyo, Japan

    K Aritaki & A Hoshika

  2. First Department of Internal Medicine, Tokyo Medical University, Tokyo, Japan

    JH Ohyashiki, A Suzuki & K Ohyashiki

  3. Department of Virology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan

    JH Ohyashiki, T Ojima, K Abe, N Shimizu & K Yamamoto

Authors
  1. K Aritaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. JH Ohyashiki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T Ojima
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K Abe
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. N Shimizu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. K Yamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. K Ohyashiki
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. A Hoshika
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Aritaki, K., Ohyashiki, J., Suzuki, A. et al. A rapid monitoring system of human herpesviruses reactivation by LightCycler in stem cell transplantation. Bone Marrow Transplant 28, 975–980 (2001). https://doi.org/10.1038/sj.bmt.1703266

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.bmt.1703266

Keywords

This article is cited by

Search

Advanced search

Quick links