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

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.

Published ahead of print on 17 January 2007.

This article has been cited by other articles:

• 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]