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The Deputy Führer of the Third Reich Rudolf Hess was captured after a controversial flight to Scotland in 1941. Hess was sentenced to life imprisonment during the Nuremberg War Crimes Trials. He was detained in Berlin's Spandau Prison under the official security designation 'Spandau #7.' Early doubts arose about the true identity of prisoner 'Spandau #7.' This evolved to a frequently espoused conspiracy theory that prisoner 'Spandau #7' was an imposter and not Rudolf Hess. After Hess's reputed 1987 suicide, the family grave became a Neo-Nazi pilgrimage site. In 2011, the grave was abandoned and the family remains cremated. Here we report the forensic DNA analysis of the only known extant DNA sample from prisoner 'Spandau #7' and a match to the Hess male line, thereby refuting the Doppelgänger Theory.

When bodies are highly decomposed and identification based on morphologic features is no longer possible, short tandem repeat (STR) genotyping has been proved as a convenient and reliable alternative. However, at very advanced decomposition stages postmortal tissue putrefaction processes can affect microsatellite instability and therefore not all types of post mortem tissues are still equally suitable for STR genotyping and subsequent forensic identification. In many cases bone or dental material is used but here reprocessing is very elaborate and expensive. Thus, multiple studies have focused on the eligibility of soft tissue and organs where DNA extraction is a lot easier and faster. In this study different cases were evaluated in which a successful forensic identification was only possible with DNA material extracted from specific tissues. By doing so we aim to compare differential suitability of soft tissues for DNA analysis of highly decomposed bodies. This was achieved by a systematic classification of decomposed bodies and the analysis of multiple organs to evaluate the qualification of soft tissue for DNA profiling in respect to the degree of decomposition. In summary, we detected differences in the suitability of various soft tissues for DNA-based identification. Cerebral matter turned out to be one of the most useful targets in highly decomposed bodies.

In forensic genotyping the gender specific marker Amelogenin is a valuable tool for gender determination. This marker is therefore standard in all commercial forensic genetic kits. However, gonosomale aberrations, deletions or primer binding site mutations can cause problems in the correct interpretation of the marker Amelogenin and consequently a valid sex determination. Misleading results can be obtained based on incorrect interpretation of the phenotype or genetic intersexuality even though the genotyping based on Amelogenin itself was performed and evaluated correctly. In our routine casework we observed six cases with discrepancies regarding the biological and the “legal” gender. Further investigation of these cases revealed varying reasons for these discrepancies. But not all cases could be completely resolved and clarified.

With the new 6-dye AmpFISTR(®) Yfiler(®) Plus amplification kit (Thermo Fisher Scientific, Waltham, MA, USA) a set of 25 Y-chromosomal short tandem repeat loci (Y-STRs), including seven rapidly mutating Y-STRs (RM Y-STRs), is now available for forensic DNA typing. In this study we present our validation data for the AmpFISTR(®) Yfiler(®) Plus amplification kit and show the results of Y-chromosomal typing of 425 unrelated male individuals from two Austrian populations (Salzburg and Upper Austria) with the AmpFISTR(®) Yfiler(®) Plus amplification kit. Forensic parameters were calculated and compared for four Y-STR marker sets. We also typed five brother pairs to evaluate the power of discrimination for related individuals. The AmpFISTR(®) Yfiler(®) Plus (Yfiler Plus) kit appeared to be unimpaired by typical inhibitors such as hematin and humic acid or by large amounts of female components. An upgrade of analyzed markers resulted in increased discrimination capacity that is crucial for forensic trace analysis.

Forensic DNA analyses have become more and more sensitive in the past years. With the ability to generate DNA profiles even from minute amounts of cellular material also the possibility to detect DNA on trace material that originates from persons not linked to the crime event, such as crime scene investigators, increases. The contamination of crime scene samples can lead to false positive results and misinterpretation that can cause deceptive investigations. In this work we continue a study of 2010 that compared the number of detected contamination incidents that were caused in the pre-analytical phase of forensic DNA analysis with the number of crime scene samples analyzed by our laboratory. Within the past 17 years we were able to detect a total of 347 contamination incidents caused by police officers in approximately 46,000 trace samples to their origin (0.75%). Additionally we demonstrate the usefulness of reference profile databases that contain DNA profiles of police officers to detect contamination incidents of trace material.

A badly decomposed body required identification by means of DNA analysis. A brother and sister of the deceased were available as reference subjects. Although investigation of Y-chromosomal markers established an exclusion condition, autosomal markers suggested a positive identification. In order to increase the reliability of the tests, X-chromosomal markers were also investigated. This analysis showed the body to have an XXY genotype (Klinefelter's syndrome). A number of hypotheses were assessed using biostatistical methods, ultimately resulting in a definite identification. The special aspect of Klinefelter's syndrome proved highly useful for biostatistical analysis.

With the development of more sensitive DNA amplification and STR typing kits, the chance of detecting minute contamination of trace material increases. At the same time, possible ways of detecting unwanted contamination of crime scene samples were also enhanced. To estimate the contamination rate of biological traces nationwide, we continued a study from 2010 that compares the number of analyzed crime scene samples with the number of detected contamination incidents within our laboratory. The results of this study illustrate the importance of reference profiles, especially of crime scene investigators, and the advantage of database assisted profile alignment in cases of mixtures.

Due to its high reliability, DNA-typing is the method of preference in the field of osseous human remains identification. Nevertheless, contaminations from various sources have been shown to be inherent to the system, especially if the DNA-yield of samples under investigation is expected to be at a low level. For this reason a special focus has to be put on sampling procedures and contamination control in order to prevent from false results. In this study we present an illustrative case report followed by particular recommendations for taking samples from osseous human remains.

Y-STR markers are a valuable tool in the analysis of biological traces in which a mixture of male and female trace material is to be expected. It is possible to generate a Y-chromosome DNA profile, even if all the prior sperm tests are negative and no sign of any male component is found in amelogenin. In 38 of a total of 239 sexual offences a perpetrator trace was identified solely using Y-STR analysis. Based on these findings, the Austrian National DNA Database was expanded to include Y-STRs in 2012 with the primary objective to identify serial sexual offences.

The introduction of five "new European standard loci" (D1S1656, D2S441, D10S1248, D12S391 and D22S1045) in forensic DNA analysis called for the development of new Multiplex PCR kits. One of the aims was to combine the widespread SGM+ marker combination with the new markers to be amplified in a single kit. In addition, SE33 was also to be included in the new kits. We present practical experiences that have been made since the introduction of PowerPlex ESI 17 and AmpflSTR NGM SElect in our laboratory.