Direct-to-PCR tissue preservation for DNA profiling

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• 作者：Amy Sorensen ; Clare Berry ; David Bruce…
• 刊名：International Journal of Legal Medicine
• 出版年：2016
• 出版时间：May 2016
• 年：2016
• 卷：130
• 期：3
• 页码：607-613
• 全文大小：530 KB
• 刊物类别：Medicine
• 刊物主题：Medicine & Public Health
  Forensic Medicine
  Medical Law
  Medicine/Public Health, general
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1437-1596
• 卷排序：130

文摘

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.KeywordsDisaster victim identification (DVI)Mass disasterTissue preservationDNA profileDirect PCRPCR inhibition