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Rapid quantitation of cytomegalovirus DNA in whole blood by a new molecular assay based on automated sample preparation and real-time PCR

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Abstract

Quantitation of cytomegalovirus (CMV) DNA in whole blood samples has gained significance in the routine diagnostic laboratory. In this study, the analytical performance of the artus™ CMV RG PCR kit in conjunction with automated sample preparation on the new QIAsymphony™ SP instrument was evaluated. Clinically referred samples were tested and results compared to those obtained with the routinely used molecular test system. Accuracy testing showed results to be within ±0.2 log10 unit of the expected panel results. The assay was linear (R = 0.9972) from a lower quantification limit of 148 copies/ml to 1.3 × 107 copies/ml. The between-day imprecision CV was 8–63%, and the within-run imprecision CV was 16–61%. When 100 clinically referred samples were tested, results obtained with the new test system showed an acceptable concordance with those obtained with the routinely used easyMAG™ sample preparation and CMV HHV6,7,8 R-gene™ test system. Discrepant results were found with low-titer samples containing CMV DNA concentrations under the lower limit of quantification or within half a log unit above. The time to results was comparable for both test systems. The QIAsymphony™ sample preparation and artus™ CMV RG PCR test system allows for a rapid, sensitive, precise, and accurate high-throughput quantitation of CMV DNA in whole blood in the routine diagnostic laboratory.

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Acknowledgments

We gratefully acknowledge Evelyn Stelzl for fruitful discussions and Egon Marth for continuous support. Reagents required for this study were supported in part by Qiagen.

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Authors and Affiliations

  1. Clinical Institute of Medical and Chemical Laboratory Diagnostics, University Hospital Graz, Graz, Austria

    Reinhard B. Raggam

  2. Research Unit Molecular Diagnostics, Center of Applied Biomedicine, IHME, Medical University of Graz, Universitaetsplatz 4, 8010, Graz, Austria

    Michael Bozic & Harald H. Kessler

  3. Section of Infectious Diseases, Division of Pulmonology, Department of Internal Medicine, Medical University of Graz, Graz, Austria

    Helmut J. F. Salzer

  4. R&D Department, Qiagen GmbH, Hilden, Germany

    Sandra Hammerschmidt & Cordula Homberg

  5. Institute of Laboratory Diagnostics, Kaiser-Franz-Josef-Hospital, Vienna, Austria

    Katharina Ruzicka

Authors
  1. Reinhard B. Raggam
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  2. Michael Bozic
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  3. Helmut J. F. Salzer
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  4. Sandra Hammerschmidt
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  5. Cordula Homberg
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  6. Katharina Ruzicka
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  7. Harald H. Kessler
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Corresponding author

Correspondence to Harald H. Kessler.

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Raggam, R.B., Bozic, M., Salzer, H.J.F. et al. Rapid quantitation of cytomegalovirus DNA in whole blood by a new molecular assay based on automated sample preparation and real-time PCR. Med Microbiol Immunol 199, 311–316 (2010). https://doi.org/10.1007/s00430-010-0164-z

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  • DOI: https://doi.org/10.1007/s00430-010-0164-z

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