title: Preservation and rapid purification of DNA from decomposing human tissue samples.
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.
authors.affiliation: Department of Forensic Science, College of Criminal Justice, Sam Houston State University, Huntsville, TX, 77341 USA. Electronic address: aes049@shsu.edu.
authors.forename: Amy
authors.initials: A
authors.lastname: Sorensen
authors.affiliation: Department of Forensic Science, College of Criminal Justice, Sam Houston State University, Huntsville, TX, 77341 USA. Electronic address: Elizabeth.Rahman@austintexas.gov.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Rahman
authors.affiliation: Department of Forensic Science, College of Criminal Justice, Sam Houston State University, Huntsville, TX, 77341 USA. Electronic address: cassandra.canela@dps.texas.gov.
authors.forename: Cassandra
authors.initials: C
authors.lastname: Canela
authors.affiliation: Department of Forensic Science, College of Criminal Justice, Sam Houston State University, Huntsville, TX, 77341 USA. Electronic address: dag006@shsu.edu.
authors.forename: David
authors.initials: D
authors.lastname: Gangitano
authors.affiliation: Department of Forensic Science, College of Criminal Justice, Sam Houston State University, Huntsville, TX, 77341 USA. Electronic address: sxh039@shsu.edu.
authors.forename: Sheree
authors.initials: S
authors.lastname: Hughes-Stamm
mesh_terms: D004247:DNA / Q000032:analysis*; D016172:DNA Fingerprinting; D005404:Fixatives / Q000737:chemistry*; D005838:Genotype; D006358:Hot Temperature; D006801:Humans; D006813:Humidity; D018482:Muscle, Skeletal / Q000473:pathology; D016133:Polymerase Chain Reaction; D011180:Postmortem Changes; D012867:Skin / Q000473:pathology; D013048:Specimen Handling; D014021:Tissue Preservation / Q000379:methods*
chemical_list: D005404:Fixatives; D004247:DNA
keywords: Disaster victim identification (DVI); Forensic science; STR typing; Tissue preservation