Operationalisation of the ForenSeq® Kintelligence Kit for Australian unidentified and missing persons casework

Area of Science:

• Forensic Genetics
• Human Identification
• Molecular Biology

Background:

• Single nucleotide polymorphism (SNP) genotyping is vital for human remains identification, aiding in estimating biological sex, biogeographical ancestry (BGA), externally visible characteristics (EVCs), identity, uniparental lineage, and kinship.
• Forensic laboratories require robust tools for analyzing diverse and often degraded human samples.

Purpose of the Study:

• To validate and operationalize the ForenSeq® Kintelligence Kit for coronial casework in human remains identification.
• To assess the kit's performance in generating genetic intelligence, including accuracy in predicting biological sex, EVCs, BGA, and kinship relationships.

Main Methods:

• Internal validation of the ForenSeq® Kintelligence Kit according to SWGDAM guidelines, focusing on typical casework sample types.
• Optimization of a high-throughput library preparation and sequencing workflow, including development of analytical thresholds for low DNA input samples.
• Comparison of genetic intelligence generated by the kit with self-declared donor data and assessment of kinship accuracy using GEDmatch PRO™.

Main Results:

• The kit produced high-quality SNP profiles from 0.1-1.0 ng DNA with high repeatability and reproducibility.
• Prediction accuracies included biological sex (95%), hair color (58%), eye color (74%), and BGA inferences (74% consistent).
• The kit accurately detected (97%) and classified (90%) genetic relationships from first to fifth degree.

Conclusions:

• The ForenSeq® Kintelligence Kit is a validated, robust, and reliable genetic intelligence tool for the Australian Federal Police National DNA Program for Unidentified and Missing Persons.
• The kit enables high-throughput analysis of challenging forensic samples for human identification, utilizing existing laboratory instrumentation.
• This SNP genotyping assay significantly enhances capabilities in investigating unidentified and missing persons cases.