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Journal of Clinical Microbiology, August 2000, p. 2929-2932, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Real-Time PCR for Diagnosis and Follow-Up of Toxoplasma Reactivation after Allogeneic Stem Cell Transplantation Using Fluorescence Resonance Energy Transfer Hybridization Probes

J.-M. Costa,1 C. Pautas,2 P. Ernault,1 F. Foulet,3 C. Cordonnier,2 and S. Bretagne3,*

Laboratoire de Biologie Moléculaire, Hôpital Américain, BP 109, 92202, Neuilly,1 and Service d'Hématologie Clinique, Hôpital H. Mondor-APHP,2 and Laboratoire de Parasitologie, Hôpital H. Mondor-APHP et Université Paris XII,3 94010, Créteil, France

Received 5 January 2000/Returned for modification 5 April 2000/Accepted 19 May 2000

Toxoplasma reactivation is a life-threatening complication of allogeneic stem cell transplantation. A poor prognosis is probably linked to a difficult diagnosis, based on the detection of evidence of parasites in tissue. We developed a real-time PCR test using fluorescence resonance energy transfer hybridization probes to detect and quantify Toxoplasma gondii DNA in serum. This PCR test gave reproducible quantitative results over a dynamic range of from 0.75 × 106 to 0.75 parasites per PCR mixture. Serial samples from four patients with toxoplasma reactivation were evaluated. Three patients had several consecutive PCR-positive samples which corresponded to <= 0.75 parasites. These three patients became PCR negative during trimethoprim-sulfamethoxazole therapy but never developed clinically apparent toxoplasmosis. In contrast, one patient had an increasing PCR signal, from 1 to 396 parasites in 12 days, and developed cerebral symptoms. The parasite count decreased to 5 parasites in 3 days after pyrimethamine-clindamycin treatment. Real-time quantitative PCR is useful for diagnosis and follow-up of toxoplasma reactivation.

* Corresponding author. Mailing address: Laboratoire de Parasitologie-Mycologie, Hôpital Henri Mondor, 51 avenue du Général DeLattre de Tassigny, 94010, Créteil, Cedex, France. Phone: 33-1-49-81-36-41. Fax: 33-1-49-81-36-01. E-mail: bretagne{at}univ-paris12.fr.

Journal of Clinical Microbiology, August 2000, p. 2929-2932, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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