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Journal of Clinical Microbiology, March 2007, p. 1042-1044, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.01109-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Real-Time PCR Diagnostics Failure Caused by Nucleotide Variability within Exon 4 of the Human Cytomegalovirus Major Immediate-Early Gene{triangledown}

Martina Lengerova,1* Zdenek Racil,2 Pavlina Volfova,1 Jana Lochmanova,1 Jitka Berkovcova,1 Dana Dvorakova,1 Jiri Vorlicek,2 and Jiri Mayer2

Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematooncology, University Hospital Brno, Brno, Czech Republic,1 Department of Internal Medicine-Hematooncology, University Hospital Brno, Brno, Czech Republic2

Received 30 May 2006/ Returned for modification 17 July 2006/ Accepted 4 January 2007

Here we report how variability in the human cytomegalovirus genome sequence may seriously affect the outcome of its molecular diagnosis. A real-time quantitative PCR assay targeting the major immediate-early gene failed to detect the viral load in some, but not all, clinical samples from hematooncological patients. By amplification and sequencing the DNA across the regions targeted by this assay we found a number of nucleotide substitutions which accounted for decreased primer/probe binding. This decreased the sensitivity of the assay up to 1,000-fold.

* Corresponding author. Mailing address: Center of Molecular Biology and Gene Therapy, University Hospital, Cernopolni 9, CZ-613 00 Brno, Czech Republic. Phone and fax: 420-532 234 623. E-mail: mlengerova{at}fnbrno.cz.

{triangledown} Published ahead of print on 17 January 2007.

Journal of Clinical Microbiology, March 2007, p. 1042-1044, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.01109-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hoffman, N. G., Cook, L., Atienza, E. E., Limaye, A. P., Jerome, K. R. (2008). Marked Variability of BK Virus Load Measurement Using Quantitative Real-Time PCR among Commonly Used Assays. J. Clin. Microbiol. 46: 2671-2680 [Abstract] [Full Text]  
  • Leruez-Ville, M., Ducroux, A., Rouzioux, C. (2008). Exon 4 of the Human Cytomegalovirus (CMV) Major Immediate-Early Gene as a Target for CMV Real-Time PCR. J. Clin. Microbiol. 46: 1571-1572 [Full Text]  


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