ArticleJournal of clinical microbiology2005
Sequence diversity in the glycoprotein B gene complicates real-time PCR assays for detection and quantification of cytomegalovirus.
Article in Journal of clinical microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- False-negative CMV PCR results due to viral sequence variation: a diagnostic pitfall with the potential for serious consequences.ASM case reports · 2026Article
- Viral infections associated with oral cancers and diseases in the context of HIV: a workshop report.Oral diseases · 2016Article
- Impact of genomic sequence variability on quantitative PCR assays for diagnosis of polyomavirus BK infection.Journal of clinical microbiology · 2011Article
- International standards and reference materials for quantitative molecular infectious disease testing.The Journal of molecular diagnostics : JMD · 2010Review
- High genotypic diversity and a novel variant of human cytomegalovirus revealed by combined UL33/UL55 genotyping with broad-range PCR.Virology journal · 2009Article
- Biologic diversity of polyomavirus BK genomic sequences: Implications for molecular diagnostic laboratories.Journal of medical virology · 2008Article
- Marked variability of BK virus load measurement using quantitative real-time PCR among commonly used assays.Journal of clinical microbiology · 2008Article
- Active CMV disease does not always correlate with viral load detection.Bone marrow transplantation · 2007Article
- Real-time PCR diagnostics failure caused by nucleotide variability within exon 4 of the human cytomegalovirus major immediate-early gene.Journal of clinical microbiology · 2007Article
- LightUp probes in clinical diagnostics.Molecular aspects of medicineReview
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Authors and funding
5 authors.
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No grant is acknowledged in the PubMed record.
Abstract
Real-time quantitative PCR systems (Q-PCR) for the rapid detection and quantification of microorganisms in clinical specimens employ oligodeoxyribonucleotide primers and probes for specificity, which makes them vulnerable to false negatives caused by sequence diversity in the template. Schaade et al. (J. Clin. Microbiol. 39:3809, 2001) reported a sequence variant (C630T) in the cytomegalovirus (CMV) glycoprotein B (gB) gene that, although detectable in their Q-PCR assay, could not be accurately quantified. In an effort to evaluate the impact of CMV sequence variants in our patient population by use of a similar Q-PCR assay, we surveyed 54 isolates of CMV, each from a different patient. We detected evidence for the C630T variant in 4 of 54 (7.4%) patients. Furthermore, isolates from two additional patients were completely negative in the test. Sequencing of these false-negative isolates revealed multiple mutations within the probe hybridization sites. A Q-PCR that targeted the CMV polymerase gene instead of gB detected all 54 isolates. We suggest that Q-PCR assays for viral load be rigorously tested on large panels of viral isolates to assess the impact of sequence diversity on detection as well as quantification.
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