Comparison of two nucleic acid extraction and testing systems for HCMV-DNA detection and quantitation on whole blood specimens from transplant patients

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• 作者：Cristina Costa ; Samantha Mantovani ; Cinzia. Balloco ; Francesca Sidoti ; Fabrizio Fop ; Rossana Cavallo
• 关键词：Human cytomegalovirus ; Nucleic acid ; Extraction ; Quantitative PCR ; Efficiency ; Variability
• 刊名：Journal of Virological Methods
• 出版年：2013
• 出版时间：November, 2013
• 年：2013
• 卷：193
• 期：2
• 页码：579-582
• 全文大小：425 K

文摘

Quantitative detection of human cytomegalovirus (HCMV) DNA on whole blood is currently the primary choice for virological monitoring in transplant patients and for determining the appropriate antiviral strategy, however specific issues of variability remain in terms of extraction methods, amplification efficiency, and variability. This study compared the performance characteristics of two nucleic acid extraction and testing systems for HCMV-DNA quantitation, the artus^? CMV QS-RGQ kit, associated with a fully automated DNA extraction and assay set up by Qiagen (system 1) and the Q-CMV Real Time Complete kit by Nanogen, associated with a semiautomated nucleic acid extraction system by Biom¨¦rieux (system 2) in 189 specimens from transplant patients and 10 from 2012 HCMV Quality Control for Molecular Diagnostics (QCMD). The two systems exhibited a 80.4 % concordance. Differences between the two systems were within ¡À1 log₁₀ copies/ml of the averaged log₁₀ results for 88.9 % of the tested specimens. For all qualitatively discordant specimens, mean viral load was ¡Ü3 log₁₀ copies/ml. Considering viral load measurement, system 1 gave earlier positives that system 2, with a 14.8 % of specimens resulted positive at low viral loads with system 1 and negative with system 2. In QCMD specimens, difference was below 0.7 log₁₀ copies/ml for both the systems.

In conclusion, the two systems provided reliable and comparable results. Some specific performance characteristic and automation could be taken into account in terms of less hands of time, fewer errors and reliability.