P189 – PCR artefacts in forensic STR typing using GlobalFiler™ IQC

11:01

Thursday August 20^th

Station 06

Duration: 12 minutes

03. Human identification

Christel Enroth

STR-based DNA analysis remains the primary approach for human identification in forensics. The use of DNA profiles in kinship analysis, criminal casework and the identification of missing persons rely on the ability to consistently produce DNA profiles of high quality.

The context in which samples for analysis have been collected may vary greatly and factors such as degradation, low copy number or multiple contributors complicate the interpretation of DNA profiles.

As the amplification and CE techniques are advanced and offer enhanced detection and increased sensitivity, with lowered input requirements and degenerative primers, the occurrence of artefacts is in turn increased as well, as shown by a growing body of evidence.

Non-human artefacts can often be distinguished based on characteristics owing to their presumed non-STR based origin. The characteristics include but are not limited to: The apparent lack of stutter (1); An unusual allelic balance compared to the other contributor components in the sample or replica samples (2); Artefacts found in samples taken from swaps of the same origin (rocks, windows sills or vagina) have the same migration length (3). Artefacts that occur in samples extracted by differential extraction appear to be extracted with different efficiency, which could be explained by their underlining microbial origin (4). Artefacts very often disappear when the sample is run with a different kit (5). An artefact can be accompanied by co-artefacts with pairs or triplets of consistent migration length combinations (6).

While the artefacts can be recognized and to some extent accounted for during data analysis, their presence complicates DNA profile comparisons. The potential presence of artefacts should be a subject of attention, especially for semi-automated processes such as Number of Contributors estimation (NOC) and probabilistic genotyping, where the presence of non-human artefacts may interfere and potentially lead to erroneous contributor estimations and/or genotype distributions.

We consider the awareness of potential artefacts in STR profiles of great importance. The validation and exchange of knowledge and information regarding non-human STRs within the scientific community is key.

We have collected information about artefacts encountered in our lab and we have tried to validate them, in part with the use of alternative kits. Here we present a selection of artefacts collected using the GlobalFiler™ IQC PCR kit, which to our knowledge has not been shared previously. We show examples of how the artefacts present themselves in DNA profiles and how they can affect the outcome of probabilistic genotyping.

Author

Christel Enroth (Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark)

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P189 – PCR artefacts in forensic STR typing using GlobalFiler™ IQC

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