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Does zero really mean nothing?—first experiences with the new PowerQuantTM system in comparison to established real-time quantification kits

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Abstract

DNA quantification is an important step in the molecular genetic analysis of a forensic sample, hopefully providing reliable data on DNA content for a subsequent generation of reproducible STR profiles for identification. For several years, this quantification has usually been done by real-time PCR protocols and meanwhile a variety of assays are commercially available from different companies. The newest one is the PowerQuantTM assay by Promega Inc. which is advertised with the promise that a determined DNA concentration of 0 ng/μl in a forensic sample guarantees the impossibility to achieve true STR results, thus allowing to exclude such samples from STR analysis to save time and money. Thus, the goal of this study was to thoroughly verify the quantification step with regard to its suitability as a screening method. We have evaluated the precision and reliability of four different real-time PCR quantification assays by systematically testing DNA dilutions and forensic samples with various DNA contents. Subsequently, each sample was subjected to the Powerplex® ESX 17 fast kit to determine a reliable cutoff level for exclusion of definitely negative samples from STR analysis. An accurate quantification of different cell line DNA dilutions was not possible with any kit. However, at least the PowerQuantTM assay provided suitable data analyzing forensic samples, whereas in other systems up to 46 % of negative samples still displayed reliable STR analysis results. All in all, the PowerQuantTM assay represents a big step forward, but the evaluation of real-time PCR quantification results has still to be done with great care.

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Authors and Affiliations

  1. Institute of Legal Medicine, University Hospital Essen, Hufelandstr. 55, D-45122, Essen, Germany

    Micaela Poetsch, Helen Konrad, Janine Helmus & Thomas Bajanowski

  2. ForScix GmbH, Hamburg, Germany

    Nicole von Wurmb-Schwark

Authors
  1. Micaela Poetsch
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  2. Helen Konrad
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  3. Janine Helmus
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  4. Thomas Bajanowski
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  5. Nicole von Wurmb-Schwark
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Correspondence to Micaela Poetsch.

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All samples were obtained after informed consent and with approval of the Medical Ethics Committee at the University of Duisburg-Essen in accordance with the Declaration of Helsinki and national laws.

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Poetsch, M., Konrad, H., Helmus, J. et al. Does zero really mean nothing?—first experiences with the new PowerQuantTM system in comparison to established real-time quantification kits. Int J Legal Med 130, 935–940 (2016). https://doi.org/10.1007/s00414-016-1352-1

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  • DOI: https://doi.org/10.1007/s00414-016-1352-1

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