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Direct-to-PCR tissue preservation for DNA profiling.

Amy Sorensen1, Clare Berry1, David Bruce1

  • 1National Centre for Forensic Studies, Faculty of Education, Science, Technology & Mathematics (ESTeM), University of Canberra, Canberra, Australia.

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Summary

This study demonstrates that direct DNA profiling from tissue samples is possible without extraction, using specialized preservatives. This method streamlines disaster victim identification (DVI) and ensures successful DNA typing even after long-term storage.

Keywords:
DNA profileDirect PCRDisaster victim identification (DVI)Mass disasterPCR inhibitionTissue preservation

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Area of Science:

  • Forensic Science
  • Molecular Biology
  • Biochemistry

Background:

  • Disaster victim identification (DVI) faces challenges in remote locations due to limited access to mortuary facilities and refrigeration.
  • Effective DNA sampling and preservation are crucial for successful DNA profiling and timely repatriation of remains.
  • Streamlining the DVI process by eliminating DNA extraction steps could significantly improve efficiency.

Purpose of the Study:

  • To investigate the feasibility of obtaining DNA profiles directly from preserved human tissues without prior DNA extraction.
  • To evaluate the efficacy of different preservative solutions in maintaining DNA integrity for profiling.
  • To assess the impact of long-term storage on DNA profiling success.

Main Methods:

  • Fresh and decomposing human muscle and skin tissues were preserved using four different solutions: two custom (DMSO/EDTA and TENT buffer) and two proprietary (DNAgard and Tissue Stabilising Kit).
  • Aliquots of the preservative solutions containing tissue were directly added to polymerase chain reaction (PCR) for DNA profiling.
  • DNA profiles were generated using PowerPlex 21 and GlobalFiler systems.
  • Preserved samples were stored at 35 °C for up to 28 days, and aliquots stored at -80 °C for 4 years were also tested.

Main Results:

  • Full DNA profiles were successfully obtained from both fresh and decomposed tissues using all four preservatives after 28 days of preservation at 35 °C.
  • DNA profiles were also successfully generated from tissue samples stored in preservative solutions at -80 °C for 4 years.
  • Long-term storage at -80 °C appeared to reduce PCR inhibition, potentially enhancing DNA typing success.

Conclusions:

  • Direct DNA profiling from preserved tissues is a viable method for disaster victim identification (DVI).
  • The tested preservative solutions effectively maintain DNA quality for profiling, even under challenging conditions and long-term storage.
  • Eliminating DNA extraction simplifies the DVI process, enabling faster results and repatriation.