Next-generation Sequencing (NGS) Feasibility and Guidance Study for Forensic DNA.

As submitted by the proposer: In our increasingly interconnected and complex world, the ability to uniquely identify individuals, generate investigative leads, and attribute trace-level evidence are becoming essential for the intelligence community, national security, and public safety. As this reality emerges, new technologies continue to evolve that enable more precise identification and characterization of individuals. Battelle proposes to conduct a 19-month applied research project in partnership with forensic DNA scientists from academia; a renowned government reference laboratory; and established city, county, state, and federal law enforcement laboratories. To objectively guide the use and eventual adoption of next-generation sequencing (NGS) in the broader DNA forensics community, the Battelle Team will conduct NGS feasibility and validation testing. The proposed study will be segmented into Performance Testing (Phase 1) and Interlaboratory Testing (Phase 2). During Phase 1, technical leads from all participating institutions will be involved in identifying key requirements and performance evaluation criteria for NGS systems (hardware, reagents, software) that would be critical for acceptance of this new technology by the forensic DNA community. DNA samples provided by the National Institute of Standards and Technology (NIST) will be sequenced on the Illumina MiSeq FGx platform or the LifeTech Ion PGM Sequencing System for initial performance testing. In addition, commercially available short tandem repeat (STR), single nucleotide polymorphism (SNP), Y-STR, and mitochondrial DNA reagent kits will be used, and data will be analyzed using Battelle's proprietary BioForgeTM software or NGS instrument-specific software. Battelle will present the performance findings to NIJ for down selection. Phase 2 interlaboratory testing will be managed by Battelle and performed by The Pennsylvania State University (PSU); NIST; the Armed Forces DNA Identification Laboratory (AFDIL); the Bureau of Alcohol, Tobacco, Firearms and Explosives (ATFE); the California Department of Justice (CAL DOJ); the Philadelphia Police Department (PPD); and the Harris County Institute of Forensic Sciences (HCIFS). To ensure quality and reduce variability, Battelle will conduct onsite assessments of individual laboratories. Data generated by participating laboratories will be transferred to Battelle for additional analyses, and then for eventual archiving and storage. Battelle will lead NIJ briefings on the results, publish results in peer-reviewed scientific journals, and disseminate the information via instructional webinars and other educational materials. The Battelle Team views the outcomes of the study as instrumental to advancing the technical knowledge of forensic DNA scientists, ultimately facilitating their contribution to the development of strategic roadmaps for technology acceptance and transition. ca/ncf