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Journal of Clinical Microbiology, December 2001, p. 4362-4369, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4362-4369.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Quantification of Human Cytomegalovirus DNA in Bone Marrow Transplant Recipients by Real-Time PCR

Frank Griscelli,1,* Michel Barrois,2 Sylvie Chauvin,1 Stephane Lastere,1 Dominique Bellet,3 and Jean-Henri Bourhis4

Service de Microbiologie,1 Service de Génétique Moléculaire,2 Service d'Immunologie,3 and Service d'Hématologie Oncologique,4 Institut Gustave-Roussy, 94805 Villejuif Cedex, France

Received 24 May 2001/Returned for modification 22 August 2001/Accepted 18 September 2001

A real-time PCR assay was developed to quantify human cytomegalovirus (CMV) DNA in peripheral blood leukocytes (PBLs) of bone marrow transplantation patients. Unlike other teams, we quantified CMV and the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene using a plasmid containing both sequences as an external standard. Tenfold serial dilutions of this plasmid yielded overlapping standard curves that allowed the quantification of CMV and GAPDH gene copies in an efficient and accurate manner. Sequential blood samples (164 specimens) were collected from 16 patients. PBLs were tested by the pp65 antigenemia assay and quantitative CMV and GAPDH gene PCRs. CMV DNA was detected by PCR in 13 patients a mean of 15 days prior to the appearance of antigenemia. The administration of anti-CMV drugs led to a rapid decrease in the numbers of viral copies and positive nuclei. Real-time PCR assay results correlated with those of the CMV pp65 antigenemia assay (P < 0.00001). The TaqMan assay may be a useful tool for rapid quantification of CMV infection and for monitoring of CMV reactivation in bone marrow transplantation recipients.

* Corresponding author. Mailing address: Laboratoire de Microbiologie Médicale, Institut Gustave-Roussy, 39 rue Camille Desmoulins, 94805 Villejuif Cedex, France. Phone: (33-1) 42 11 51 93. Fax: (33-1) 42 11 53 13. E-mail: grisceli{at}igr.fr.

Journal of Clinical Microbiology, December 2001, p. 4362-4369, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4362-4369.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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