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Journal of Clinical Microbiology, September 2000, p. 3249-3253, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

False-Positive Results of Plasma PCR for Cytomegalovirus DNA due to Delayed Sample Preparation

Peter Schäfer,1,* Werner Tenschert,2 Matthias Schröter,1 Kai Gutensohn,3 and Rainer Laufs1

Institut für Medizinische Mikrobiologie und Immunologie,1 Klinik und Poliklinik für Urologie,2 and Abteilung für Transfusionsmedizin,3 Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany

Received 28 December 1999/Returned for modification 25 April 2000/Accepted 22 June 2000

Positive results by cytomegalovirus (CMV) PCR of plasma are considered predictive of active CMV infection in kidney allograft recipients. To assess whether contamination with leukocyte-derived CMV DNA can distort the results, aliquots of whole-blood samples from 60 CMV immunoglobulin G-positive patients with leukocyte CMV DNAemia were stored for up to 24 h at room temperature (RT) and at 4°C before plasma preparation. Native and ultrafiltered plasma samples were tested by CMV and beta -globin PCRs. Among 30 latently infected patients (negative for CMV pp65 antigens), low baseline rates (10%) and levels (median number of copies, 10 [per 10 µl]) of CMV plasma DNAemia in native plasma samples increased significantly over time (after 4 h at RT, 37% [P < 0.001]; median number of copies, 45 [P < 0.001]). Similar effects were found during storage at 4°C. Ultrafiltration reduced the levels of CMV plasma DNAemia, but by 6 h of storage the levels were significantly elevated as well. CMV and beta -globin DNA kinetics in plasma were parallel. In contrast, 30 actively infected patients (pp65 positive) had high baseline rates (87% in native samples) and levels (median number of copies, 75) of CMV plasma DNAemia. No significant effects of storage or ultrafiltration and no concordance with beta -globin DNA kinetics were seen. In conclusion, delayed preparation of plasma samples bears a significant risk of false-positive CMV PCR results, probably due to leukocyte lysis. This has important implications in the clinical setting and for PCR standardization.

* Corresponding author. Mailing address: Institut für Medizinische Mikrobiologie und Immunologie, Universitätsklinikum Hamburg-Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany. Phone: (49 40) 42803-3157. Fax: (49 40) 42803-4881. E-mail: pschaefe{at}uke.uni-hamburg.de.

Journal of Clinical Microbiology, September 2000, p. 3249-3253, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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