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J. Clin. Microbiol. doi:10.1128/JCM.01403-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Detection of Cytomegalovirus DNA in EDTA Whole Blood Samples: Evaluation of the Quantitative artus® CMV LC PCR Kit in Conjunction with Automated Sample Preparation

Birgit D. A. Michelin, Ita Hadisejdi, Michael Bozic, Maja Grahovac, Markus Hess, Blaenka Grahovac, Egon Marth, and Harald H. Kessler^*

Institute of Hygiene, Medical University of Graz, Graz, Austria; Department of Pathology, Faculty of Medicine, University of Rijeka, Rijeka, Croatia; Qiagen Hamburg GmbH, Research and Development, Hamburg, Germany

^* To whom correspondence should be addressed. Email: harald.kessler{at}meduni-graz.at.

	Abstract

Whole blood has been found to be a reliable matrix for detection and quantitation of cytomegalovirus DNA. In this study, the performance of the artus® CMV LC PCR Kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation was evaluated. Accuracy, linearity, analytical sensitivity, inter- and intra-assay variations were determined. A total of 102 clinical EDTA whole blood samples were investigated and results were compared with those obtained by the IVD (in vitro diagnostics)/CE-labeled CMV HHV6,7,8 R-gene Quantification kit. When accuracy of the new kit was tested, seven of eight results were found to be within ± 0.5 log₁₀ unit of the expected panel results. Determination of linearity resulted in a quasilinear curve over more than 5 log units. The lower limit of detection of the assay was determined to be 139 copies/ml in EDTA whole blood. The interassay variation ranged from 15 to 58%, and the intra-assay variation ranged from 7 to 35%. When clinical samples were tested and compared with the routinely used IVD/CE-labeled assay, 53 samples tested positive and 13 samples negative with both of the assays. One sample was found to be positive with the artus® CMV LC PCR Kit only and 35 samples tested positive with the routinely used assay only. The majority of discrepant results were found with low-titer samples. In conclusion, the artus® CMV LC PCR Kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation may be suitable for detection and quantitation of CMV DNA in EDTA whole blood in the routine low-throughput laboratory; however, low-positives may be missed by this assay.