Direct-to-PCR tissue preservation for DNA profiling.

Sorensen, A; Berry, C; Bruce, D; Gahan, ME; Hughes-Stamm, S; McNevin, D

Direct-to-PCR tissue preservation for DNA profiling.

Sorensen, A Berry, C Bruce, D Gahan, ME Hughes-Stamm, S McNevin, D

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Publisher:: SPRINGER
Publication Type:: Journal Article
Citation:: Int J Legal Med, 2016, 130, (3), pp. 607-613
Issue Date:: 2016-05

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Field	Value	Language
dc.contributor.author	Sorensen, A
dc.contributor.author	Berry, C
dc.contributor.author	Bruce, D
dc.contributor.author	Gahan, ME
dc.contributor.author	Hughes-Stamm, S
dc.contributor.author	McNevin, D https://orcid.org/0000-0003-1665-3367
dc.date.accessioned	2022-01-31T22:32:46Z
dc.date.available	2015-10-22
dc.date.available	2022-01-31T22:32:46Z
dc.date.issued	2016-05
dc.identifier.citation	Int J Legal Med, 2016, 130, (3), pp. 607-613
dc.identifier.issn	0937-9827
dc.identifier.issn	1437-1596
dc.identifier.uri	http://hdl.handle.net/10453/154001
dc.description.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.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	SPRINGER
dc.relation.ispartof	Int J Legal Med
dc.relation.isbasedon	10.1007/s00414-015-1286-z
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0399 Other Chemical Sciences, 0699 Other Biological Sciences, 1103 Clinical Sciences
dc.subject.classification	Legal & Forensic Medicine
dc.subject.mesh	Cryoprotective Agents
dc.subject.mesh	DNA
dc.subject.mesh	DNA Fingerprinting
dc.subject.mesh	Dimethyl Sulfoxide
dc.subject.mesh	Edetic Acid
dc.subject.mesh	Genotype
dc.subject.mesh	Humans
dc.subject.mesh	Microsatellite Repeats
dc.subject.mesh	Muscle, Skeletal
dc.subject.mesh	Polymerase Chain Reaction
dc.subject.mesh	Skin
dc.subject.mesh	Specimen Handling
dc.subject.mesh	Tissue Preservation
dc.subject.mesh	Muscle, Skeletal
dc.subject.mesh	Skin
dc.subject.mesh	Humans
dc.subject.mesh	Edetic Acid
dc.subject.mesh	Dimethyl Sulfoxide
dc.subject.mesh	DNA
dc.subject.mesh	Cryoprotective Agents
dc.subject.mesh	Specimen Handling
dc.subject.mesh	Tissue Preservation
dc.subject.mesh	DNA Fingerprinting
dc.subject.mesh	Polymerase Chain Reaction
dc.subject.mesh	Microsatellite Repeats
dc.subject.mesh	Genotype
dc.title	Direct-to-PCR tissue preservation for DNA profiling.
dc.type	Journal Article
utslib.citation.volume	130
utslib.location.activity	Germany
utslib.for	0399 Other Chemical Sciences
utslib.for	0699 Other Biological Sciences
utslib.for	1103 Clinical Sciences
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - CFS - Centre for Forensic Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Mathematical and Physical Sciences
utslib.copyright.status	closed_access	*
dc.date.updated	2022-01-31T22:32:45Z
pubs.issue	3
pubs.publication-status	Published
pubs.volume	130
utslib.citation.issue	3

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.

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http://hdl.handle.net/10453/154001