Developmental validation of the ForenSeq® Kintelligence kit, MiSeq FGx® sequencing system and ForenSeq Universal...

Area of Science:

• Forensic Genomics
• Genetic Genealogy
• Next-Generation Sequencing (NGS)

Background:

• Forensic Investigative Genetic Genealogy (FIGG) is an emerging field combining DNA analysis with genealogical research.
• FIGG aids in identifying unidentified human remains and generating investigative leads in criminal cases.
• Current methods require sensitive and high-throughput technologies for effective DNA evidence analysis.

Purpose of the Study:

• To describe the design and validation of the ForenSeq Kintelligence Kit and associated workflow.
• To evaluate the kit's performance for forensic DNA evidence analysis and investigative genetic genealogy.
• To enable the operational transfer of FIGG to forensic laboratories.

Main Methods:

• Development of the ForenSeq Kintelligence multiplex assay (10,230 SNPs).
• Utilizing the MiSeq FGx Sequencing System and ForenSeq Universal Analysis Software.
• Developmental validation following SWGDAM guidelines for an end-to-end workflow.

Main Results:

• The ForenSeq Kintelligence Kit provides more genetic information from challenging samples compared to existing assays.
• Higher marker density and shorter amplicon sizes (97.8% <150 bp) enhance data retrieval.
• The assay demonstrated robustness across a wide range of sample quantities and qualities.

Conclusions:

• The ForenSeq Kintelligence Kit and software are fit for purpose for forensic investigative genetic genealogy.
• This workflow facilitates the integration of FIGG into operational forensic laboratory settings.
• The technology enhances the identification of human remains and supports criminal investigations.