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A novel approach for rapid cell assessment to estimate DNA recovery from human bone tissue

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Abstract

We report on the use of a DNA staining dye to locate and record nucleated osteocytes and other bone-related cells within sections of archived formalin-fixed and paraffin-embedded human tibia from which informative DNA profiles were obtained. Eleven of these archived tibia samples were sectioned at a thickness of 5 µm. Diamond™ Nucleic Acid Dye was applied to the sections and cells within the matrix of the bone fluoresced so that their location and number of cells could be photographed. DNA was isolated from these 11 samples using a standard extraction process and the yields were quantified by real-time PCR. Complete STR profiles were generated from ten bone extracts where low-level inhibition was recorded with an incomplete STR profile obtained from one sample with higher inhibition. The stained image of this sample showed that few cells were present. There was a significant relationship between the number of DD-stained cells and the number of alleles obtained (p < 0.05). Staining cells to determine the prevalence of bone cell nuclei allows a triage of samples prior to any subsequent DNA profiling.

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Acknowledgements

A special thanks to Mr. Adnan Mulaibrahimovic and Ms. Pat Vilimas for their assistance to TNL in performing histological samples.

Funding

This research was funded by the Attorney General’s Department through Forensic Science SA and the Ross Vining Memorial Research Fund.

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Authors and Affiliations

  1. College of Science and Engineering, Flinders University, Flinders, SA, 5042, Australia

    Thien Ngoc Le, Oliva Handt, Julianne Henry & Adrian Linacre

  2. Discipline of Orthopaedics and Trauma, Adelaide Medical School, The University of Adelaide, Adelaide, Australia

    Dzenita Muratovic

  3. Forensic Science SA, PO Box 2790, Adelaide, SA, 5001, Australia

    Oliva Handt & Julianne Henry

Authors
  1. Thien Ngoc Le
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  2. Dzenita Muratovic
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  4. Julianne Henry
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Contributions

TNL designed and performed the experiments. DM provided the samples, assisted with experimental design and practical advice. AL assisted with manuscript drafting, provided experimental advice, facilitated access the laboratory and funded this study. JMH assisted with the manuscript drafting and provided experimental advice. OH assisted with the manuscript drafting and provided experimental advice.

Corresponding author

Correspondence to Adrian Linacre.

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Ethical approval

This study was approved by the Human Research Ethics Committees of the Royal Adelaide Hospital and The University of Adelaide, South Australia. Bone samples used in this study were collected with informed written consent from patients who had undergone knee arthroplasty surgery. All samples were anonymized with the exception of age and sex.

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None.

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Le, T.N., Dzenita Muratovic, Handt, O. et al. A novel approach for rapid cell assessment to estimate DNA recovery from human bone tissue. Forensic Sci Med Pathol 17, 649–659 (2021). https://doi.org/10.1007/s12024-021-00428-3

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