Highlights
- •We evaluated five DNA preservatives (modified TENT, DESS, LST, RNAlater and DNAgard®) on decomposing tissues stored in hot and humid conditions.
- •A marked decrease in DNA quantity and quality was observed in all samples after the bodies entered the bloat stage of decomposition.
- •Similar amounts of DNA were retrieved from skin and muscle samples over time, but slightly more complete STR profiles were obtained from muscle tissue.
- •Although higher amounts of DNA were recovered from tissue samples, the completeness of STR profiles from “free DNA” in solution was comparable.
- •DNAgard® and the modified TENT buffer were the most successful preservatives based on STR success from “free DNA” in solution.
Abstract
One of the key features to be considered in a mass disaster is victim identification.
However, the recovery and identification of human remains are sometimes complicated
by harsh environmental conditions, limited facilities, loss of electricity and lack
of refrigeration. If human remains cannot be collected, stored, or identified immediately,
bodies decompose and DNA degrades making genotyping more difficult and ultimately
decreasing DNA profiling success. In order to prevent further DNA damage and degradation
after collection, tissue preservatives may be used.
The goal of this study was to evaluate three customized (modified TENT, DESS, LST)
and two commercial DNA preservatives (RNAlater and DNAgard®) on fresh and decomposed human skin and muscle samples stored in hot
(35 °C) and humid (60–70% relative humidity) conditions for up to three months. Skin and
muscle samples were harvested from the thigh of three human cadavers placed outdoors
for up to two weeks. In addition, the possibility of purifying DNA directly from the
preservative solutions (“free DNA”) was investigated in order to eliminate lengthy
tissue digestion processes and increase throughput.
The efficiency of each preservative was evaluated based on the quantity of DNA recovered
from both the “free DNA” in solution and the tissue sample itself in conjunction with
the quality and completeness of downstream STR profiles. As expected, DNA quantity
and STR success decreased with time of decomposition. However, a marked decrease in
DNA quantity and STR quality was observed in all samples after the bodies entered
the bloat stage (approximately six days of decomposition in this study). Similar amounts
of DNA were retrieved from skin and muscle samples over time, but slightly more complete
STR profiles were obtained from muscle tissue. Although higher amounts of DNA were
recovered from tissue samples than from the surrounding preservative, the average
number of reportable alleles from the “free DNA” was comparable. Overall, DNAgard®
and the modified TENT buffer were the most successful tissue preservatives tested
in this study based on STR profile success from “free DNA” in solution when decomposing
tissues were stored for up to three months in hot, humid conditions.
Keywords
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Article info
Publication history
Published online: May 23, 2016
Accepted:
May 23,
2016
Received in revised form:
May 4,
2016
Received:
October 25,
2015
Identification
Copyright
© 2016 Elsevier Ireland Ltd. All rights reserved.
