Eva Simeoni

A new multiplex-PCR comprising autosomal and y-specific STRs and mitochondrial DNA to analyze highly degraded material

The analysis of short tandem repeats is one of the most powerful tools in forensic genetics. Forensic practice sometimes requires the individualization of samples that may contain only highly degraded nuclear DNA, mitochondrial DNA or PCR inhibitors that hamper DNA amplification. We designed a new multiplex PCR with reduced size amplicons (<200 bp), providing a double sex determination (amelogenin plus two Y-STRs), the detection of two autosomal markers and the amplification of mitochondrial specific fragments from the hypervariable region I (HVI). Additionally, a quality sensor was developed to check for the presence of any PCR inhibitors. The new multiplex PCR shows a reproducible detection threshold down to 25 pg and gives signals even out of highly degraded materials. All signals are reproducible and reliable as it could be shown in comparison to results from commercially available STR multiplex-PCRs. In no case DNA fragments were detectable using any other assay when the quality sensor was not detectable. There was a good correlation between detection of mitochondrial specific fragments in the multiplex-PCR and success of subsequent sequencing of HVI region. The same could be shown for STR analysis: Most samples successfully analyzed in our PCR yielded at least a partial STR profile using a commercial STR kit. We present an assay that allows an easy, reliable, and cost efficient evaluation of DNA sample quality combined with a first rough sample individualization and sex determination suitable for forensic purposes. This assay may help the forensic lab personnel to decide on further sample processing.

Genetic investigation of modern burned corpses

Abstract Burning of corpses is a well-known and widely spread funeral procedure that has been performed for a long time in many cultures. Nowadays more and more corpses are burned in cremations and buried in urns, often for practical and financial reasons. Usually cremation takes place as temperatures of over 1000 °C for more than 70 min, destroying the corpse and only leaving severely burned teeth and some fragments of larger bones. In some scientific, criminal or civil cases even after cremation, there is the need of genetic investigations for identification or paternity testing. Here we present a systematic genetic investigation of 10 corpses that were burned in a crematory. After DNA extraction of the remains, the presence of human nuclear and mitochondrial DNA was tested by a highly sensitive Duplex PCR and quantified via real time PCR; genetic typing was done using the AmpFlSTR Identifiler kit.

Biostatistical basis of individualisation and segregation analysis using the multilocus DNA probe MZ 1.3: Results of a collaborative study

A collaborative study using the multilocus minisatellite DNA probe MZ 1.3 was carried out to investigate segregation information, mutation rate, DNA fragment frequencies as well as band sharing characteristics. The fingerprint patterns of 393 children as well as 694 unrelated individuals were analysed after digestion of DNA with the restriction enzyme HinfI. A mutation rate of 1% per meiosis or 0.04% per band was found with a mean number of 26 bands/individual. It was shown that maternal and paternal fragments are inherited in equal proportions. Population frequencies of restriction fragments demonstrated a distribution with increasing frequencies in the small fragment size range below 10 kb as well as the absence of very common or very rare fragments. Our data can be used to calculate simple exclusion probabilities based on the number of non-maternal bands in the child.

An Illicit Love Affair During the Third Reich: Who is My Grandfather?*

Abstract:  More than 60 years after an illicit love affair had occurred between Erika H, wife of a Wehrmacht soldier, and a Polish slave worker during World War II, we could clarify the blood relationships of her daughter Uta. When Erika H had become pregnant both of the men could have fathered the child. Erika H was found guilty of fraternization and imprisoned at Ravensbrück concentration camp. She gave birth to Uta and died there in 1944. Uta survived the war as did Erika’s husband Gustav, who accepted Uta as his child. Blood samples from family members were taken and DNA extracted. A panel of 16 short tandem repeat (STR) loci were amplified and separated by capillary electrophoresis and the likelihoods calculated using the MLINK software. The combined genotypes yielded a cumulative likelihood ratio of over 200,000 against paternity of Gustav H. This case serves to illustrate the utility of STR profiles for complex deficiency kinship analysis.

Identification of blood group substances of fingerprints on the murder weapon by means of the adhesive strip method

Earlier investigations by ISHIYAMA, PROKOP and GOHLER as well as KEIL and PAPSDORF (1977) have shown that it is possible to demonstrate blood group substances on fingerprints. We reported elsewhere (SIMEONI, GRUNER and BURKHARDT 1984) on the application of the absorption elution method in the determination of the main blood group from fingerprints in excretors and nonexcretors of blood group substances.