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Optimization strategies for DNA investigations of highly degraded corpses

Disaster victim identification

Optimierter Workflow für DNA-Analysen an hochgradig postmortal veränderten Leichen

Identifikation von Katastrophenopfern

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Abstract

Background

The successful post-mortem identification of highly degraded corpses and mass disaster victims using forensic molecular genetic analyses is a challenging task and particularly influenced by the efficacy and evaluation of standard examinations in the laboratory workflow. In many cases only highly degraded soft tissue is present, so that only bones, teeth or nails are available for DNA analysis. A lack of expertise in bone preparation for DNA typing in conjunction with the choice of time-consuming methods often leads to large delays during victim identification.

Objectives

The present study was carried out to provide an alternative method aimed at efficient mechanical grinding of bone and molecular genetic analyses in conjunction with time-minimizing strategies.

Material and methods

The sample batch consisted of diaphysis sections from the femurs of 18 corpses with post-mortem intervals (PMIs) ranging from 1 week to 5.5 years. Time saving was achieved by optimizing sample preparation and genotyping processes (i.e. DNA purification, quantification and STR analysis) in this study.

Results

In comparison with other commonly used procedures, which require 1 to 2 weeks for complete analysis, it was possible to obtain DNA identification profiles in only 2.5 days. Semi-automated sub-processes for certain steps of the complete workflow allowed minimal staffing with a high sample throughput (50 bones per week).

Conclusion

The present workflow supplements existing standard procedures in the laboratory especially in cases where a large number of low quality samples are to be tested and DNA results are urgently needed, e.g. in disaster victim identification (DVI).

Zusammenfassung

Hintergrund

Die erfolgreiche Identifizierung Verstorbener mit fortgeschrittenen postmortalen Veränderungen, z. B. nach Massenkatastrophen, mithilfe forensischer molekulargenetischer Analysen stellt immer noch eine Herausforderung für die Laborroutine dar. Der Analyseerfolg wird maßgeblich durch existierende Prüfstandards im Arbeitsablauf und die Laborlogistik beeinflusst. In vielen Fällen ist das Weichgewebe so verändert, dass nur Knochen, Zähne bzw. Nägel für die DNA-Analyse zur Verfügung stehen. Fehlendes Know-how im Bereich der Vorbereitung für die DNA-Analytik an Knochenmaterial in Verbindung mit der Auswahl zeitaufwendiger Methoden führt oft zu einer enormen zeitlichen Verzögerung bei der Identifizierung.

Ziel

Vor diesem Hintergrund wird innerhalb der Studie ein alternativer methodischer Workflow für die DNA-Typisierung vorgestellt.

Material und Methoden

Untersucht wurden Diaphysenabschnitte der Femora von 18 Verstorbenen, die unterschiedlich stark ausgeprägte Erscheinungen fortgeschrittener Leichenveränderungen aufwiesen. Das postmortale Intervall lag zwischen einer Woche und 5,5 Jahren. Eine Zeitersparnis im Analyseprozess wurde durch Optimierung der Probenaufbereitungstechnik und einzelner DNA-Analyseschritte (DNA-Isolation, Quantifizierung, STR-Analyse) erreicht.

Ergebnisse

Durch die Optimierung einzelner Prozessschritte und Anwendung neuartiger, sensitiver DNA-Analyse Kits war es möglich, das Zeitfenster für einen vollständigen Analyseprozess von bisher benötigten 1–2 Wochen, auf 2,5 Tage zu minimieren. Der optimierte Workflow mit für manche Teilprozesse semiautomatisierten Prozessen ermöglicht einen hohen Probendurchlauf (50 Knochen pro Woche) bei minimalem personellem Aufwand.

Zusammenfassung

Der Workflow ergänzt bestehende Standardprozeduren im forensischen Labor, vor allem wenn eine große Anzahl qualitativ minderwertiger Proben unter hohem Zeitdruck zu untersuchen ist, beispielsweise zur Opferidentifizierung nach Massenkatastrophen.

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Authors and Affiliations

  1. Institute of Legal Medicine, Johannisallee, 04103, Leipzig, Germany

    A.-M. Pflugbeil, M. Harthun, J. Edelmann, J. Dreßler & K. Thiele

  2. Forensic Science Investigation Lab (FoSIL), University of Applied Sciences, Technikumplatz 17, 09648, Mittweida, Germany

    D. Labudde

Authors
  1. A.-M. Pflugbeil
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  2. M. Harthun
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  3. D. Labudde
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  4. J. Edelmann
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  5. J. Dreßler
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  6. K. Thiele
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Correspondence to A.-M. Pflugbeil.

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Conflict of interest

A.M. Pflugbeil, M. Harthun, D. Labudde, J. Edelmann, J. Dreßler and K. Thiele declare that there are no conflicts of interest. All analyses on samples described in this study were carried out in accordance with the Helsinki Declaration of 1964 (in its current, revised form, 2013).

The accompanying manuscript does not include studies on humans or animals.

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Pflugbeil, AM., Harthun, M., Labudde, D. et al. Optimization strategies for DNA investigations of highly degraded corpses. Rechtsmedizin 25, 268–273 (2015). https://doi.org/10.1007/s00194-015-0042-5

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  • DOI: https://doi.org/10.1007/s00194-015-0042-5

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