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.

  • Original Article
  • Published:

Infections Post Transplant

Diagnosis of toxoplasmosis in bone marrow transplant recipients: comparison of PCR-based results and immunohistochemistry

Abstract

Toxoplasmosis in bone marrow transplant recipients is a rare but serious complication and if untreated, almost uniformly fatal. The diagnosis, however, remains difficult. We therefore compared serial determination of antibody titers specific for T. gondii before and after transplantation, serial PCR for T. gondii DNA in serum, PCR and nested PCR for T. gondii DNA in various tissues, conventional histology and immunohistochemistry for detection of parasites in three patients with autopsy-confirmed toxoplasmosis after bone marrow transplantation. Immunohistochemistry demonstrated the presence of parasites in 13 out of 20 organs investigated (65%), whereas PCR detected T. gondii-specific DNA in 15 out of 20 organs (75%). Immunohistochemistry revealed concordant results to PCR data in 60% of the specimens. With the use of a nested PCR protocol, eight out of nine samples (89%) were positive for T. gondii-specific DNA. The combination of both methods detected the presence of parasites in 90% of the specimens. Serial PCR in serum did not yield positive results. Neither PCR nor immunohistochemistry was able to detect parasites in all organs investigated, but both methods together improved sensitivity to 90% and consequently, should be used jointly to maximize diagnostic precision. Bone Marrow Transplantation (2000) 25, 1257–1262.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Similar content being viewed by others

References

  1. Beaman MH, McCabe RE, Wong S-Y et al. Toxoplasma gondii. In: Mandell GL, Bennett JE, Dolin R (eds) Principles and Practice of Infectious Diseases Churchill Livingstone: New York 1995 2455–2475

    Google Scholar 

  2. Chandrasekar PH, Momin F, the Bone Marrow Transplant Team . Disseminated toxoplasmosis in marrow recipients: a report of three cases and a review of the literature Bone Marrow Transplant 1997 19: 685–689

    Article  CAS  Google Scholar 

  3. Slavin MA, Meyers JD, Remington JS et al. Toxoplasma gondii infection in marrow transplant recipients: a 20 year experience Bone Marrow Transplant 1994 13: 549–557

    CAS  Google Scholar 

  4. Derouin F, Gluckman E, Beauvais B et al. Toxoplasma infection after human allogeneic bone marrow transplantation: clinical and serological study of 80 patients Bone Marrow Transplant 1986 1: 67–73

    CAS  Google Scholar 

  5. Israelski DM, Remington JS . Toxoplasmosis in the non-AIDS immunocompromised host. In: Remington JS, Schwartz M (eds) Current Clinical Topics in Infectious Diseases Blackwell Scientific: Boston 1993 322–356

    Google Scholar 

  6. Shepp DH, Hackman RC, Conley FK et al. Toxoplasma gondii reactivation identified by detection of parasitemia in tissue culture Ann Intern Med 1985 103: 218–221

    Article  CAS  Google Scholar 

  7. Derouin F, Devergie A, Auber P et al. Toxoplasmosis in bone marrow-transplant recipients: report of seven cases and review Clin Infect Dis 1992 15: 267–270

    Article  CAS  Google Scholar 

  8. Bottone EJ . Diagnosis of acute pulmonary toxoplasmosis by visualization of invasive and intracellular tachyzoites in Giemsa-stained smears of bronchoalveolar lavage fluid J Clin Microbiol 1991 29: 2626–2627

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Bretagne S, Costa JM, Kuentz M et al. Late toxoplasmosis evidenced by PCR in a marrow transplant recipient Bone Marrow Transplant 1995 15: 809–811

    CAS  Google Scholar 

  10. Roth A, Roth B, Höffken G et al. Application of the polymerase chain reaction in the diagnosis of pulmonary toxoplasmosis in immunocompromised patients Eur J Clin Microbiol Infect Dis 1992 11: 1177–1181

    Article  CAS  Google Scholar 

  11. Bundesgesundheitsamt Sabin-Feldman-Test (SFT) zum Nachweis von Antikörpern gegen Toxoplasma gondii (Routinemethode) Bundesgesundheitsblatt 1989 32: 553–555

  12. Goelz SE, Hamilton SR, Vogelstein B . Purification of DNA from formaldehyde fixed and paraffin embedded human tissue Biochem Biophys Res Commun 1985 130: 118–126

    Article  CAS  Google Scholar 

  13. Krüger D, Weise W, Bug C et al. Investigations on selected red cell concentrates for Toxoplasma and Toxoplasma–DNA Infusionsther Transfusionsmed 1997 24: 82–85

    Google Scholar 

  14. Khalifa KES, Roth A, Roth B et al. Value of PCR for evaluating occurrence of parasitemia in immunocompromised patients with cerebral and extracerebral toxoplasmosis J Clin Microbiol 1994 32: 2813–2819

    Google Scholar 

  15. Ellis JT, Luton K, Baverstock PR et al. Phylogenetic relationships between Toxoplasma and Sarcocystis deduced from a comparison of 18S rDNA sequences Parasitology 1995 110: 521–528

    Article  CAS  Google Scholar 

  16. Cazenave J, Cheyrou A, Begueret J . Improvement of rDNA-based PCR assay to detect Toxoplasma Prenat Diag 1993 13: 543

    Article  CAS  Google Scholar 

  17. Heurkens AHM, Koelma IA, de Planque MM et al. Failure to diagnose fatal disseminated toxoplasmosis in a bone marrow transplant recipient: the possible significance of declining antibody titres J Immunol 1989 18: 283–288

    CAS  Google Scholar 

  18. Derouin F, Sarfati C, Beauvais B et al. Laboratory diagnosis of pulmonary toxoplasmosis in patients with acquired immunodeficiency syndrome J Clin Microbiol 1989 27: 1661–1663

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Crisan D, Mattson JC . Retrospective DNA analysis using fixed tissue specimens DNA Cell Biol 1993 12: 455–464

    Article  CAS  Google Scholar 

  20. Kwok S, Higuchi R . Avoiding false positives with PCR Nature 1989 339: 237–238

    Article  CAS  Google Scholar 

  21. Romero RL, Juston AC, Ballantyne J et al. The applicability of formalin-fixed and formalin fixed paraffin embedded tissues in forensic DNA analysis J Forensic Sci 1997 42: 708–714

    Article  CAS  Google Scholar 

  22. Shedlock AM, Haygood MG, Pietsch TW et al. Enhanced DNA extraction and PCR amplification of mitochondrial genes from formalin-fixed museum specimens Bio Techniques 1997 22: 394–400

    CAS  Google Scholar 

Download references

Acknowledgements

We gratefully acknowledge the technical assistance of Ms C Bug, E Kamal and G Stoffels. We also thank the nurses and physicians of our bone marrow transplant unit who excellently cared for our patients.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Held, T., Krüger, D., Switala, A. et al. Diagnosis of toxoplasmosis in bone marrow transplant recipients: comparison of PCR-based results and immunohistochemistry. Bone Marrow Transplant 25, 1257–1262 (2000). https://doi.org/10.1038/sj.bmt.1702457

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.bmt.1702457

Keywords

This article is cited by

Search

Quick links