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

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• 作者：Sabine Yerly ; Luc Perrin ; Christian Van Delden ; Sven Schaffer ; Sven Thamm ; Werner Wunderli ; Laurent Kaiser
• 关键词：Cytomegalovirus ; Real-time PCR ; Plasma CMV DNA
• 刊名：Journal of Clinical Virology
• 出版年：2007
• 出版时间：April 2007
• 年：2007
• 卷：38
• 期：4
• 页码：298-303
• 全文大小：381 K

文摘

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.