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Journal of Clinical Microbiology, January 2001, p. 251-259, Vol. 39, No. 1
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.1.251-259.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Direct Quantification of Human Cytomegalovirus Immediate-Early and Late mRNA Levels in Blood of Lung Transplant Recipients by Competitive Nucleic Acid Sequence-Based Amplification

Astrid E. Greijer,1,2 Erik A. M. Verschuuren,3 Martin C. Harmsen,3 Chantal A. J. Dekkers,1 Henriëtte M. A. Adriaanse,1 T. Hauw The,3 and Jaap M. Middeldorp1,2,*

Organon Teknika, Boxtel,1 Department of Pathology, University Hospital, Vrije Universiteit, Amsterdam,2 and Department of Clinical Immunology, University Hospital Groningen, Groningen,3 The Netherlands

Received 26 July 2000/Returned for modification 8 September 2000/Accepted 25 October 2000

The dynamics of active human cytomegalovirus (HCMV) infection was monitored by competitive nucleic acid sequence-based amplification (NASBA) assays for quantification of IE1 (UL123) and pp67 (UL65) mRNA expression levels in the blood of patients after lung transplantation. RNA was isolated from 339 samples of 13 lung transplant recipients and analyzed by the quantitative IE1 and pp67 NASBA in parallel with pp65 antigenemia and serology. Rapid increases in IE1 RNA exceeding 104 copies per 100 µl of blood were associated with active infection, whereas lower levels were suggestive for abortive, subclinical viral activity. Any positive value for pp67 RNA was indicative for active infection, and quantification of pp67 mRNA did not give additional diagnostic information. The onset of IE1-positive NASBA preceded pp67 NASBA and was earlier than the pp65 antigenemia assay, confirming previous studies with qualitative NASBA. Effective antiviral treatment was reflected by a rapid disappearance of pp67 mRNA, whereas IE1 mRNA remained detectable for longer periods. Quantification of IE1 might be relevant to monitor progression of HCMV infection but should be validated in prospective studies.

* Corresponding author. Mailing address: Department of Pathology, Academic Hospital, Vrije Universiteit, De Boelelaan 1117, 1007 MB Amsterdam, The Netherlands. Phone: 31-20-4444070. Fax: 31-20-4442964. E-mail: J.Middeldorp{at}azvu.nl.

Journal of Clinical Microbiology, January 2001, p. 251-259, Vol. 39, No. 1
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.1.251-259.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Hamprecht, K., Pfau, M., Schaller, H.-E., Jahn, G., Middeldorp, J. M., Rennekampff, H.-O. (2005). Human Cytomegalovirus Infection of a Severe-Burn Patient: Evidence for Productive Self-Limited Viral Replication in Blood and Lung. J. Clin. Microbiol. 43: 2534-2536 [Abstract] [Full Text]  


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