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Cytomegalovirus quantification in plasma by an automated real-time PCR assay

Schaffer, Sven
Thamm, Sven
Published in Journal of Clinical Virology. 2007, vol. 38, no. 4, p. 298-303
Abstract BACKGROUND: Sensitive quantitation of cytomegalovirus (CMV) DNA in blood is helpful for the diagnosis of CMV infection or reactivation and the monitoring of transplanted patients. OBJECTIVES: We compared a new PCR assay coupled with an automated extraction system (CMV real-time PCR, Abbott Molecular, Des Plaines, IL, USA) to a previously validated method (ultrasensitive Cobas Amplicor CMV DNA Monitor, Roche Molecular, Indianapolis, IN, USA). RESULTS: Using limiting dilutions of CMV DNA positive plasma, the two assays had a similar detection threshold ranging between 20 and 45 copies/ml. Coefficients of variation of CMV real-time PCR assay varied from 1 to 12% for CMV DNA levels between 10,000 and 20 copies/ml. Viral loads assessed by the two methods on 179 clinical samples showed an overall concordance of 89% and an excellent correlation (R=0.94). Discrepancies were only observed for samples with low CMV DNA levels (<300 copies/ml); 18 samples were positive by CMV real-time PCR only, and 2 samples by ultrasensitive Cobas CMV only. Values obtained by CMV real-time PCR were on average 0.4 log higher than those of ultrasensitive Cobas CMV. Successive samples of transplanted patients with evidence of CMV infection/reactivation revealed that CMV real-time PCR assay was positive earlier and for a longer period of time after treatment initiation. CONCLUSIONS: Although both assays had similar analytical performances, the CMV real-time PCR assay has the advantages of automated extraction and higher dynamic range, and shows a trend for an improved sensitivity that might impact on clinical decisions.
Keywords AdultAnalysis of VarianceCytomegalovirus/ geneticsDNA, Viral/ bloodFemaleHumansPlasma/virologyPolymerase Chain Reaction/ methodsReproducibility of ResultsSensitivity and SpecificityViral Load/ methods
PMID: 17321200
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Research group Groupe Laurent Kaiser (virologie clinique) (668)
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YERLY FERRILLO, Sabine et al. Cytomegalovirus quantification in plasma by an automated real-time PCR assay. In: Journal of Clinical Virology, 2007, vol. 38, n° 4, p. 298-303. doi: 10.1016/j.jcv.2007.01.003 https://archive-ouverte.unige.ch/unige:7705

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Deposited on : 2010-06-21

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