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Journal of Clinical Microbiology, October 2008, p. 3311-3318, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00797-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Quantification of DNA in Plasma by an Automated Real-Time PCR Assay (Cytomegalovirus PCR Kit) for Surveillance of Active Cytomegalovirus Infection and Guidance of Preemptive Therapy for Allogeneic Hematopoietic Stem Cell Transplant Recipients{triangledown}

Concepción Gimeno,1,2 Carlos Solano,3 José C. Latorre,1 Juan C. Hernández-Boluda,3 María A. Clari,1 María J. Remigia,3 Santiago Furió,3 Marisa Calabuig,3 Nuria Tormo,1 and David Navarro1,2*

Microbiology Service, Hospital Clínico Universitario,1 Department of Microbiology, School of Medicine, University of Valencia,2 Hematology and Medical Oncology Service, Hospital Clínico Universitario, Valencia, Spain3

Received 28 April 2008/ Returned for modification 26 June 2008/ Accepted 14 August 2008

The performance of a plasma real-time PCR (cytomegalovirus [CMV] PCR kit; Abbott Diagnostics) was compared with that of the antigenemia assay for the surveillance of active CMV infection in 42 allogeneic hematopoietic stem cell transplantation (Allo-SCT) recipients. A total of 1,156 samples were analyzed by the two assays. Concordance between the two assays was 82.2%. Plasma DNA levels correlated with the number of pp65-positive cells, particularly prior to the initiation of preemptive therapy. Fifty-seven episodes of active CMV infection were detected in 37 patients: 18 were defined solely by the PCR assay and four were defined on the basis of the antigenemia assay. Either a cutoff of 288 CMV DNA copies/ml or a 2.42-log10 increase of DNAemia levels between two consecutive PCR positive samples was an optimal value to discriminate between patients requiring preemptive therapy and those not requiring therapy on the basis of the antigenemia results. The real-time PCR assay allowed an earlier diagnosis of active CMV infection and was a more reliable marker of successful clearance of CMV from the blood. Analysis of the kinetics of DNAemia levels at a median of 7 days posttreatment allowed the prediction of the response to CMV therapy. Two patients developed CMV colitis. The PCR assay tested positive both before the onset of symptoms and during the disease period. The plasma real-time PCR from Abbott is more suitable than the antigenemia assay for monitoring active CMV infection in Allo-SCT recipients and may be used for guiding preemptive therapy in this clinical setting.

* Corresponding author. Mailing address: Microbiology Service, Hospital Clínico Universitario, and Department of Microbiology, School of Medicine, Av. Blasco Ibáñez 17, 46010 Valencia, Spain. Phone: 34(96)3864657. Fax: 34(96)3864173. E-mail: david.navarro{at}uv.es

{triangledown} Published ahead of print on 27 August 2008.

Journal of Clinical Microbiology, October 2008, p. 3311-3318, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00797-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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