International Journal of Hematology

, Volume 108, Issue 5, pp 535–542 | Cite as

Comparison of HHV-6 DNA detection in plasma and whole blood in allogeneic hematopoietic stem cell transplant recipients: frequent false-positive results for active HHV-6 infection using whole blood samples

  • Kuniko Takano
  • Masao OgataEmail author
  • Rie Kawano
  • Takako Satou
  • Yuko Nashimoto
  • Kuniaki Shirao
Original Article


In this prospective observational study, we compared the human herpesvirus-6 (HHV-6) DNA load in serially collected paired plasma and whole blood (WB) samples from allogeneic hematopoietic stem cell transplantation (HSCT) recipients. A total of 721 paired samples were collected from 68 recipients. The positive rate for HHV-6 DNA was 9.7 and 35.0% in plasma and WB samples, respectively (P < 0.001). The correlation of HHV-6 DNA load between plasma and WB was poor (R2 = 0.250). After reaching peak levels, HHV-6 DNA showed a delayed decrease in WB in comparison with plasma (median, 28 versus 7 days, P < 0.001). We additionally tested HHV-6 mRNA status in 95 samples from eight patients. To identify positive HHV-6 mRNA, plasma HHV-6 DNA showed 55.0% sensitivity and 100% specificity, whereas WB HHV-6 DNA showed 90.0% sensitivity and 68.0% specificity. The false-positive rate for identifying positive HHV-6 mRNA was 0% for plasma HHV-6 DNA and 32.0% for WB HHV-6 DNA. Although WB was more sensitive than plasma for detecting HHV-6 reactivation, the rates of false positivity for active HHV-6 infection were higher for WB than for plasma.


Human herpesvirus-6 Plasma Whole blood mRNA Allogeneic hematopoietic stem cell transplant 



This work was supported in part by the Japan Society for the Promotion of Science (JSPS) KAKENHI (Grant-in-Aid for Scientific Research) Grant Number JP17K09090.

Author Contributions

KT and MO were responsible for the design of the study, interpretation of the data, and writing the manuscript. KT, MO, and YN analyzed the data. Data were collected by KT, MO, and YN. Laboratory work was carried out or supervised by KT, RK, TS, and KS. All authors contributed to the revision of the manuscript and approved the final version. We thank the following individuals for their contributions to this study: Kazuhiro Kohno (Koureiren Tsurumi Hospital) and Natsumi Yoshida (Oita University Faculty of Medicine). We also thank Analisa Avila, ELS, from Edanz Group and Mike Adams for proofreading a draft of this manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Zerr DM, Corey L, Kim HW, Huang ML, Nguy L, Boeckh M. Clinical outcomes of human herpesvirus 6 reactivation after hematopoietic stem cell transplantation. Clin Infect Dis. 2005;40:932–40.CrossRefPubMedGoogle Scholar
  2. 2.
    Ogata M, Satou T, Kadota J, Saito N, Yoshida T, Okumura H, et al. Human herpesvirus 6 (HHV-6) reactivation and HHV-6 encephalitis after allogeneic hematopoietic cell transplantation: a multicenter, prospective study. Clin Infect Dis. 2013;57:671–81.CrossRefPubMedGoogle Scholar
  3. 3.
    Zerr DM, Boeckh M, Delaney C, Martin PJ, Xie H, Adler AL, et al. HHV-6 reactivation and associated sequelae after hematopoietic cell transplantation. Biol Blood Marrow Transplant. 2012;18:1700–8.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Nakayama T, Okada F, Ando Y, Honda K, Ogata M, Goto K, et al. A case of pneumonitis and encephalitis associated with human herpesvirus 6 (HHV-6) infection after bone marrow transplantation. Br J Radiol. 2010;83:e255-8.CrossRefPubMedGoogle Scholar
  5. 5.
    Hill JA, Myerson D, Sedlak RH, Jerome KR, Zerr DM. Hepatitis due to human herpesvirus 6B after hematopoietic cell transplantation and a review of the literature. Transpl Infect Dis. 2014;16:477–83.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Ljungman P, Wang FZ, Clark DA, Emery VC, Remberger M, Ringden O, et al. High levels of human herpesvirus 6 DNA in peripheral blood leucocytes are correlated to platelet engraftment and disease in allogeneic stem cell transplant patients. Br J Haematol. 2000;111:774–81.PubMedGoogle Scholar
  7. 7.
    Ueki T, Hoshi K, Hiroshima Y, Sumi M, Ichikawa N, Ogata M, et al. Analysis of five cases of human herpesvirus-6 myelitis among 121 cord blood transplantations. Int J Hematol. 2018;107:363–72.CrossRefPubMedGoogle Scholar
  8. 8.
    Ogata M, Oshima K, Ikebe T, Takano K, Kanamori H, Kondo T, et al. Clinical characteristics and outcome of human herpesvirus-6 encephalitis after allogeneic hematopoietic stem cell transplantation. Bone Marrow Transplant. 2017;52:1563–70.CrossRefPubMedGoogle Scholar
  9. 9.
    Murakami K, Kohashi S, Sakurai M, Kato J, Toyama T, Koda Y, et al. Hyponatremia associated with human herpesvirus-6 (HHV-6) encephalitis after allogeneic hematopoietic stem cell transplantation: a presentation different from HHV-6 myelitis. Int J Hematol. 2017;106:436–40.CrossRefPubMedGoogle Scholar
  10. 10.
    Zerr DM. Human herpesvirus 6 (HHV-6) disease in the setting of transplantation. Curr Opin Infect Dis. 2012;25:438–44.CrossRefPubMedGoogle Scholar
  11. 11.
    Ogata M, Fukuda T, Teshima T. Human herpesvirus-6 encephalitis after allogeneic hematopoietic cell transplantation: What we do and do not know. Bone Marrow Transplant. 2015;50:1030–6.CrossRefPubMedGoogle Scholar
  12. 12.
    Yoshikawa T, Asano Y, Ihira M, Suzuki K, Ohashi M, Suga S, et al. Human herpesvirus 6 viremia in bone marrow transplant recipients: clinical features and risk factors. J Infect Dis. 2002;185:847–53.CrossRefPubMedGoogle Scholar
  13. 13.
    Yoshikawa T, Akimoto S, Nishimura N, Ozaki T, Ihira M, Ohashi M, et al. Evaluation of active human herpesvirus 6 infection by reverse transcription-PCR. J Med Virol. 2003;70:267–72.CrossRefPubMedGoogle Scholar
  14. 14.
    Volin L, Lautenschlager I, Juvonen E, Nihtinen A, Anttila VJ, Ruutu T. Human herpesvirus 6 antigenaemia in allogeneic stem cell transplant recipients: impact on clinical course and association with other beta-herpesviruses. Br J Haematol. 2004;126:690–6.CrossRefPubMedGoogle Scholar
  15. 15.
    Zerr DM. Human herpesvirus 6 infection in hematopoietic cell transplant recipients. In: Hirsch MS, Boeckh M, section editors. Thormer AR, deputy editor. UpToDate. Accessed 30 Dec 2017.
  16. 16.
    Achour A, Boutolleau D, Slim A, Agut H, Gautheret-Dejean A. Human herpesvirus-6 (HHV-6) DNA in plasma reflects the presence of infected blood cells rather than circulating viral particles. J Clin Virol. 2007;38:280–5.CrossRefPubMedGoogle Scholar
  17. 17.
    Wada K, Kubota N, Ito Y, Yagasaki H, Kato K, Yoshikawa T, et al. Simultaneous quantification of Epstein–Barr virus, cytomegalovirus, and human herpesvirus 6 DNA in samples from transplant recipients by multiplex real-time PCR assay. J Clin Microbiol. 2007;45:1426–32.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Jeulin H, Agrinier N, Guery M, Salmon A, Clement L, Bordigoni P, et al. Human herpesvirus 6 infection after allogeneic stem cell transplantation: incidence, outcome, and factors associated with HHV-6 reactivation. Transplantation. 2013;95:1292–8.CrossRefPubMedGoogle Scholar
  19. 19.
    Inazawa N, Hori T, Yamamoto M, Hatakeyama N, Yoto Y, Nojima M, et al. HHV-6 encephalitis may complicate the early phase after allogeneic hematopoietic stem cell transplantation: detection by qualitative multiplex PCR and subsequent quantitative real-time PCR. J Med Virol. 2016;88:319–23.CrossRefPubMedGoogle Scholar
  20. 20.
    Debaugnies F, Busson L, Ferster A, Lewalle P, Azzi N, Aoun M, et al. Detection of Herpesviridae in whole blood by multiplex PCR DNA-based microarray analysis after hematopoietic stem cell transplantation. J Clin Microbiol. 2014;52:2552–6.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Ogata M, Kikuchi H, Satou T, Kawano R, Ikewaki J, Kohno K, et al. Human herpesvirus 6 DNA in plasma after allogeneic stem cell transplantation: incidence and clinical significance. J Infect Dis. 2006;193:68–79.CrossRefPubMedGoogle Scholar
  22. 22.
    Kanda Y. Investigation of the freely available easy-to-use software ‘EZR’ for medical statistics. Bone Marrow Transplant. 2013;48:452–8.CrossRefPubMedGoogle Scholar
  23. 23.
    Lazzarotto T, Chiereghin A, Piralla A, Piccirilli G, Girello A, Campanini G, et al. CMV and EBV DNA kinetics in whole blood and plasma of allogeneic hematopoietic stem cell transplant recipients. Biol Blood Marrow Transplant 2018. (e-pub ahead of print).CrossRefPubMedGoogle Scholar
  24. 24.
    Luppi M, Marasca R, Barozzi P, Ferrari S, Ceccherini-Nelli L, Batoni G, et al. Three cases of human herpesvirus-6 latent infection: integration of viral genome in peripheral blood mononuclear cell DNA. J Med Virol. 1993;40:44–52.CrossRefPubMedGoogle Scholar
  25. 25.
    Arbuckle JH, Medveczky MM, Luka J, Hadley SH, Luegmayr A, Ablashi D, et al. The latent human herpesvirus-6A genome specifically integrates in telomeres of human chromosomes in vivo and in vitro. Proc Natl Acad Sci USA. 2010;107:5563–8.CrossRefPubMedGoogle Scholar
  26. 26.
    Arbuckle JH, Pantry SN, Medveczky MM, Prichett J, Loomis KS, Ablashi D, et al. Mapping the telomere integrated genome of human herpesvirus 6A and 6B. Virology. 2013;442:3–11.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Zerr DM, Fann JR, Breiger D, Boeckh M, Adler AL, Xie H, et al. HHV-6 reactivation and its effect on delirium and cognitive functioning in hematopoietic cell transplantation recipients. Blood. 2011;117:5243–9.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Ihira M, Enomoto Y, Kawamura Y, Nakai H, Sugata K, Asano Y, et al. Development of quantitative RT-PCR assays for detection of three classes of HHV-6B gene transcripts. J Med Virol. 2012;84:1388–95.CrossRefPubMedGoogle Scholar

Copyright information

© The Japanese Society of Hematology 2018

Authors and Affiliations

  1. 1.Department of Medical Oncology and HematologyOita University Faculty of MedicineOitaJapan
  2. 2.Department of HematologyOita University HospitalOitaJapan
  3. 3.Department of Clinical LaboratoryKouseiren Tsurumi HospitalOitaJapan
  4. 4.Department of HematologyKumamoto Central HospitalKumamotoJapan

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