Impact of two different commercial DNA extraction methods on BK virus viral load


https://doi.org/10.4081/mm.2016.4825
Vol. 31 No. 1 (2016)
Submitted: 11 November 2014
Accepted: 28 January 2016
Published: 31 March 2016
DNA extraction, BKV viral load, PCR real time, transplant recipients
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Authors

  • Massimiliano Bergallo
    massimiliano.bergallo@unito.it
    Department of Public Health and Paediatric Sciences, University of Turin Medical School, Turin; Laboratory of Citoimmunodiagnostics, University Hospital of City Science and Health, Regina Margherita Children's Hospital, Turin, Italy.
  • Ilaria Galliano Department of Public Health and Paediatric Sciences, University of Turin Medical School, Turin; Laboratory of Citoimmunodiagnostics, University Hospital of City Science and Health, Regina Margherita Children's Hospital, Turin, Italy.
  • Elisa Loiacono Nephrology, Dialysis and Transplantation, University Hospital City of Science and Health, Regina Margherita Children's Hospital, Turin, Italy.
  • Francesca Ferro Laboratory of Citoimmunodiagnostics, University Hospital of City Science and Health, Regina Margherita Children's Hospital, Turin, Italy.
  • Paola Montanari Department of Public Health and Paediatric Sciences, University of Turin Medical School, Turin; Laboratory of Citoimmunodiagnostics, University Hospital of City Science and Health, Regina Margherita Children's Hospital, Turin, Italy.
  • Paolo Ravanini Laboratory of Molecular Virology, Azienda Ospedaliero-Universitaria Maggiore della Carità, Novara, Italy.
Background and aim: BK virus, a member of human polyomavirus family, is a worldwide distributed virus characterized by a seroprevalence rate of 70-90% in adult population. Monitoring of viral replication is made by evaluation of BK DNA by quantitative polymerase chain reaction. Many different methods can be applied for extraction of nucleic acid from several specimens. The aim of this study was to assess the impact of two different DNA extraction procedure on BK viral load.
Materials and methods: DNA extraction procedure including the Nuclisens easyMAG platform (bioMerieux, Marcy l’Etoile, France) and manual QIAGEN extraction (QIAGEN Hilden, Germany). BK DNA quantification was performed by Real Time TaqMan PCR using a commercial kit.
Result and discussion: The samples capacity, cost and time spent were compared for both systems. In conclusion our results demonstrate that automated nucleic acid extraction method using Nuclisense easyMAG was superior to manual protocol (QIAGEN Blood Mini kit), for the extraction of BK virus from serum and urine specimens.

Bergallo, M., Galliano, I., Loiacono, E., Ferro, F., Montanari, P., & Ravanini, P. (2016). Impact of two different commercial DNA extraction methods on BK virus viral load. Microbiologia Medica, 31(1). https://doi.org/10.4081/mm.2016.4825

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