Abstract:
In 2005, the European Network of Forensic Science Institutes (ENFSI) and the European DNA Profiling Group (EDNAP) decided on an extension of the European Standard Set (ESS) loci as a result of the signing of the Prüm Treaty. This requires European Union (EU) countries to store standardised DNA profile information in national databases, enabling these countries to share information across borders. The Biology Specialist Advisory Group (BSAG), which is a group of senior managers of the Australia and New Zealand forensic DNA laboratories have agreed to adopt the extended ESS, and subsequently designed testing and validation experiments. This research project evaluated three ESS kits according to the BSAG requirements: AmpFℓSTR® NGM SElect (from Applied Biosystems), PowerPlex® ESI 17 (from Promega), and Investigator ESSplex SE (from Qiagen). These kits target the same set of 16 loci, which includes the extended ESS. This research project aimed to evaluate the performance of the new kits and determine their suitability for implementation at the Institute of Environmental Science and Research Limited (ESR). This involved sensitivity, mixture, and concordance studies. The sensitivity study involved a range of input DNA amounts to determine the amount that will result to a good quality profile. The new kits were far more sensitive than Identifiler (the current kit used in ESR). PowerPlex ESI 17 was most sensitive, producing full profiles at 50pg DNA. The new ESS kits were also determined suitable for low copy number (LCN) DNA analysis, as full profiles were obtained from 100pg of DNA, using recommended PCR cycle numbers. A mixture study was carried out involving mixed samples of two and three contributors, at ratios prescribed by BSAG. All kits successfully amplified the different components of mixed profiles. A concordance study was also performed on five samples. Results showed that the new kits are compatible with the current system, as consistent allele calls were obtained. Furthermore, the new kits were able to analyse degraded, inhibited, and small amounts of DNA. This study showed that the new ESS kits are suitable for implementation at ESR. The information obtained from this research is preliminary, and an extensive validation study will be required for the kit ultimately chosen for implementation.