Direct to PCR tissue preservation for DNA profiling

Amy Sorensen, Clare Berry, David BRUCE, Michelle GAHAN, Sheree Hughes-Stamm, Dennis MCNEVIN

    Research output: Contribution to journalArticle

    8 Citations (Scopus)

    Abstract

    Disaster victim identification (DVI) often occurs in remote locations with extremes of temperatures and humidities. Access to mortuary facilities and refrigeration are not always available. An effective and robust DNA sampling and preservation procedure would increase the probability of successful DNA profiling and allow faster repatriation of bodies and body parts. If the act of tissue preservation also released DNA into solution, ready for polymerase chain reaction (PCR), the DVI process could be further streamlined. In this study, we explored the possibility of obtaining DNA profiles without DNA extraction, by adding aliquots of preservative solutions surrounding fresh human muscle and decomposing human muscle and skin tissue samples directly to PCR. The preservatives consisted of two custom preparations and two proprietary solutions. The custom preparations were a salt-saturated solution of dimethyl sulfoxide (DMSO) with ethylenediaminetetraacetic (EDTA) and TENT buffer (Tris, EDTA, NaCl, Tween 20). The proprietary preservatives were DNAgard (Biomatrica®) and Tissue Stabilising Kit (DNA Genotek). We obtained full PowerPlex® 21 (Promega) and GlobalFiler® (Life Technologies) DNA profiles from fresh and decomposed tissue preserved at 35 °C for up to 28 days for all four preservatives. The preservative aliquots removed from the fresh muscle tissue samples had been stored at −80 °C for 4 years, indicating that long-term archival does not diminish the probability of successful DNA typing. Rather, storage at −80 °C seems to reduce PCR inhibition
    Original languageEnglish
    Pages (from-to)607-613
    Number of pages7
    JournalInternational Journal of Legal Medicine
    Volume130
    Issue number3
    DOIs
    Publication statusPublished - May 2016

    Fingerprint

    Tissue Preservation
    DNA Fingerprinting
    Polymerase Chain Reaction
    DNA
    Disaster Victims
    Muscles
    Refrigeration
    Tromethamine
    Polysorbates
    Humidity
    Dimethyl Sulfoxide
    Human Body
    Salts
    Technology
    Skin
    Temperature

    Cite this

    Sorensen, A., Berry, C., BRUCE, D., GAHAN, M., Hughes-Stamm, S., & MCNEVIN, D. (2016). Direct to PCR tissue preservation for DNA profiling. International Journal of Legal Medicine, 130(3), 607-613. https://doi.org/10.1007/s00414-015-1286-z
    Sorensen, Amy ; Berry, Clare ; BRUCE, David ; GAHAN, Michelle ; Hughes-Stamm, Sheree ; MCNEVIN, Dennis. / Direct to PCR tissue preservation for DNA profiling. In: International Journal of Legal Medicine. 2016 ; Vol. 130, No. 3. pp. 607-613.
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    abstract = "Disaster victim identification (DVI) often occurs in remote locations with extremes of temperatures and humidities. Access to mortuary facilities and refrigeration are not always available. An effective and robust DNA sampling and preservation procedure would increase the probability of successful DNA profiling and allow faster repatriation of bodies and body parts. If the act of tissue preservation also released DNA into solution, ready for polymerase chain reaction (PCR), the DVI process could be further streamlined. In this study, we explored the possibility of obtaining DNA profiles without DNA extraction, by adding aliquots of preservative solutions surrounding fresh human muscle and decomposing human muscle and skin tissue samples directly to PCR. The preservatives consisted of two custom preparations and two proprietary solutions. The custom preparations were a salt-saturated solution of dimethyl sulfoxide (DMSO) with ethylenediaminetetraacetic (EDTA) and TENT buffer (Tris, EDTA, NaCl, Tween 20). The proprietary preservatives were DNAgard (Biomatrica{\circledR}) and Tissue Stabilising Kit (DNA Genotek). We obtained full PowerPlex{\circledR} 21 (Promega) and GlobalFiler{\circledR} (Life Technologies) DNA profiles from fresh and decomposed tissue preserved at 35 °C for up to 28 days for all four preservatives. The preservative aliquots removed from the fresh muscle tissue samples had been stored at −80 °C for 4 years, indicating that long-term archival does not diminish the probability of successful DNA typing. Rather, storage at −80 °C seems to reduce PCR inhibition",
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    Sorensen, A, Berry, C, BRUCE, D, GAHAN, M, Hughes-Stamm, S & MCNEVIN, D 2016, 'Direct to PCR tissue preservation for DNA profiling', International Journal of Legal Medicine, vol. 130, no. 3, pp. 607-613. https://doi.org/10.1007/s00414-015-1286-z

    Direct to PCR tissue preservation for DNA profiling. / Sorensen, Amy; Berry, Clare; BRUCE, David; GAHAN, Michelle; Hughes-Stamm, Sheree; MCNEVIN, Dennis.

    In: International Journal of Legal Medicine, Vol. 130, No. 3, 05.2016, p. 607-613.

    Research output: Contribution to journalArticle

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    AU - Berry, Clare

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    AU - GAHAN, Michelle

    AU - Hughes-Stamm, Sheree

    AU - MCNEVIN, Dennis

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    AB - Disaster victim identification (DVI) often occurs in remote locations with extremes of temperatures and humidities. Access to mortuary facilities and refrigeration are not always available. An effective and robust DNA sampling and preservation procedure would increase the probability of successful DNA profiling and allow faster repatriation of bodies and body parts. If the act of tissue preservation also released DNA into solution, ready for polymerase chain reaction (PCR), the DVI process could be further streamlined. In this study, we explored the possibility of obtaining DNA profiles without DNA extraction, by adding aliquots of preservative solutions surrounding fresh human muscle and decomposing human muscle and skin tissue samples directly to PCR. The preservatives consisted of two custom preparations and two proprietary solutions. The custom preparations were a salt-saturated solution of dimethyl sulfoxide (DMSO) with ethylenediaminetetraacetic (EDTA) and TENT buffer (Tris, EDTA, NaCl, Tween 20). The proprietary preservatives were DNAgard (Biomatrica®) and Tissue Stabilising Kit (DNA Genotek). We obtained full PowerPlex® 21 (Promega) and GlobalFiler® (Life Technologies) DNA profiles from fresh and decomposed tissue preserved at 35 °C for up to 28 days for all four preservatives. The preservative aliquots removed from the fresh muscle tissue samples had been stored at −80 °C for 4 years, indicating that long-term archival does not diminish the probability of successful DNA typing. Rather, storage at −80 °C seems to reduce PCR inhibition

    KW - Disaster victim identification (DVI)

    KW - Mass disaster

    KW - Tissue preservation

    KW - DNA profile

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    KW - PCR inhibition

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    M3 - Article

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    ER -

    Sorensen A, Berry C, BRUCE D, GAHAN M, Hughes-Stamm S, MCNEVIN D. Direct to PCR tissue preservation for DNA profiling. International Journal of Legal Medicine. 2016 May;130(3):607-613. https://doi.org/10.1007/s00414-015-1286-z