Biogeographical Ancestry Analyses Using the ForenSeq^TM DNA Signature Prep Kit and Multiple Prediction Tools

Area of Science:

• Forensic genetics
• Population genetics
• Human identification

Background:

• Biogeographical ancestry (BGA) inference is crucial for forensic investigations, including identifying missing persons and victims.
• The ForenSeq™ DNA Signature Prep Kit and MiSeq FGx system are utilized for BGA analysis.
• Evaluating the performance of different BGA prediction tools is essential for casework accuracy.

Purpose of the Study:

• To assess the typing performance of 56 ancestry-informative SNPs using the ForenSeq™ DNA Signature Prep Kit.
• To compare the accuracy of Universal Analysis Software (UAS), FROG-kb, and GenoGeographer for BGA prediction.
• To determine the optimal DNA input amount for reliable BGA inference.

Main Methods:

• Analysis of 177 samples using 56 ancestry-informative SNPs with the ForenSeq™ DNA Signature Prep Kit on a MiSeq FGx system.
• Comparison of BGA prediction accuracy across three software tools (UAS, FROG-kb, GenoGeographer) using diverse sample populations.
• Evaluation of DNA input quantity effects on SNP typing and BGA profile completeness.

Main Results:

• The ForenSeq™ kit demonstrated high sensitivity, yielding complete SNP profiles with as low as 250pg DNA input.
• FROG-kb and GenoGeographer achieved high accuracy rates for both European (99.6%, 91.8%) and non-European (95.4%, 90.9%) samples.
• Universal Analysis Software (UAS) showed high accuracy for Europeans (96.0%) but significantly lower accuracy for non-Europeans (40.9%).
• No tested tool accurately predicted individuals with co-ancestry.

Conclusions:

• While the ForenSeq™ kit is sensitive, a minimum of 1ng DNA input is recommended to avoid low read depth and SNP drop-out.
• FROG-kb and GenoGeographer are reliable tools for BGA inference in forensic casework, particularly for diverse populations.
• Employing multiple BGA prediction tools enhances the overall accuracy and reliability of biogeographical ancestry estimation in forensic science.