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Advances in DNA sequencing technology have enabled unprecedented access to study the genetic profile of organisms, revolutionized modern medicine, and enhanced diagnostics capabilities. When warfighters deploy to austere environments under potential threats from biological warfare agents (BWAs), rapid and timely threat detection is essential to prescribe appropriate medical countermeasures along with broader Joint Force health protections.

The Defense Threat Reduction Agency’s (DTRA) Chemical and Biological Technologies Department in its role as the Joint Science and Technology Office (JSTO) for Chemical and Biological Defense is investing in a project with the Pacific Northwest National Laboratory (PNNL) to adapt a portable, commercial-off-the-shelf nanopore sequencing technology for rapid detection and diagnostics of proteins at the point of care.

Point-of-care diagnostics is essential for immediate medical response time, reducing mortality, and preserving mission effectiveness. Unfortunately, presently available diagnostic tools in this area rely primarily on traditional immunology and polymerase chain reaction technologies, often compounding the detection of BWA-specific targets and being dependent on sample concentration. Current research and development efforts are focused on using new technologies, including biosensors, nanotechnologies, and high-sensitivity molecular assays, to improve on the limitations of traditional diagnostics assays.

The same way technological advancements have led to smaller, cheaper, and more powerful computers, DNA sequencers have come a long way over the past 20 years—from large room-sized instruments to small hand-held sequencers. One such miniature technology is nanopore-based sequencing, which introduces compact, lightweight, and highly specific sequencing platforms. These features provide a unique opportunity to develop portable sequencing capabilities for biological detection and diagnostics in the field.

A hand-held platform that can detect proteins and sequence DNA would be suitable for field-forward environments where traditional medical laboratory equipment cannot go. This expanded capability to detect a broad range of biological threat agents in the field would be invaluable to the Joint Force, especially involving new or emerging threats.

The nanopore DNA sequencer is a laboratory device to determine the exact sequence of nucleotides, or bases, in a DNA molecule. This effort conducted by DTRA JSTO and PNNL uses the Oxford Nanopore Technologies MinION system to also monitor protein activity in clinical and environmental samples. The researchers are adapting validated benchtop assays to the MinION platform and are using MinION sequencing probes to serve as molecular substrates for BWAs that target host nucleic acids and proteins. Researchers successfully adapted probes against four targets with high sensitivity in laboratory samples and are ready to begin testing clinical and environmental samples such as blood, saliva, soil, and water. While there are several more studies to complete, this data supports nanopore technologies as a viable option for portable point-of-care diagnostics in field-forward settings.

DTRA JSTO continues to leverage cutting-edge technologies to provide warfighters with new diagnostics capabilities and to enhance troop preparedness. This hand-held platform will also provide a multiplexed solution for several biological threat agents that can readily adapt to emerging biological threats.

POC: Patricia McMahon, Ph.D., patricia.t.mcmahon.civ@mail.mil