TECHNOLOGY AREA(S): Chemical/Biological Defense
OBJECTIVE: Leverage phage-based technologies to develop fieldable assays and demonstrate the long-term stability of these assays.
DESCRIPTION: Technologies that enable biological detection and presumptive identification with low operational burden are needed as future Warfighter capabilities. Lateral flow immunoassays (LFIs) have remained the go-to technology for approximately 20 years despite a plethora of lab-based techniques that vastly outstrip LFIs in critical qualities such as sensitivity and specificity. This persistence of LFIs demonstrates the overwhelming importance of ease of use and low operational burden to technology adoption. While huge investments in pushing lab-based techniques towards ruggedization and simple operation are resulting in advances, platform technologies that start from a position of low operational burden and expand capabilities beyond LFIs are attractive. Meanwhile, interest in phage for a range of applications has significantly increased in recent years. Department of Defense (DoD)-relevant applications include bacterial detection, identification, decontamination, and treatment, especially of antimicrobial resistant strains. Phage offers high host specificity, built-in replication, abundance in nature, and ease of production, amongst other properties. Indeed, modified phages have been demonstrated in the lab as a highly sensitive and specific method to detect biological warfare agents such as Bacillus anthracis and Yersinia pestis. Research into the stability of phage after lyophilization spanning several decades has shown mixed results for different phage and for different preparation methods; yet, significant success suggests that phage could present an excellent approach to fielded biological detection. This topic seeks innovative development of phage-based detection and identification technologies that are highly fieldable. More specifically, ideal assays would be simple to operate, inexpensive, disposable, and require little or no equipment to analyze results; however, they must also continue to enhance the sensitivity and specificity of the assay. Determining the technical merit of using phage as the main component of fieldable assays, by determining a method for stabilization, would help drive forward the current metrics for detection and identification. While an LFI format is not specifically requested, proposed approaches should use the success of this format as a template; especially the ability to demonstrate operability in austere environments is highly encouraged.
PHASE I: Proof-of-concept will require the production of at least one specific assay that incorporates the use of a stable phage as a main component of the final design. This assay should address the ability to operate in austere environments, while continuing to be simple to operate, inexpensive (ie. less than $100/test, with a clear path towards cost reduction), disposable, and require little or no equipment to analyze results. For demonstration purposes in Phase I, detection limits are not as essential as stability of the assay components.
PHASE II: The offeror will develop functioning assays with improved limits of detection for more than one target of specific interest to the DoD. Detection of actual threat agents is encouraged but not necessarily required. However, demonstration of enhanced stability capabilities above what was demonstrated in Phase I, such as resistance to environmental fluctuations (ie. storage temperatures up to 30ºC, with up to 50ºC being optimal, and operational temperatures robust up to a 5ºC variation from ideal conditions set by the offeror) regardless of the target is also expected. At the conclusion of Phase II, the assays developed should be able to be tested for both reproducibility and accuracy of results at several storage and operational temperatures. Furthermore, suitable testing partners should be identified for threat agent testing, as work with many specific agents is highly restricted.
PHASE III DUAL USE APPLICATIONS: Efforts in both Phase II and Phase III should be clearly directed towards transition to field use. Potential limitations in sample preparation requirements, sensitivity, robustness, etc. should be clearly indicated. Avenues to overcome these limitations in potential Phase III work should be outlined. Potential products using the same technology that are not specific to DoD needs may involve different limitations (e.g. robustness to austere operations), and these separate limitations should also be outlined. Inexpensive, disposable, ruggedized detection of biological threat materials or other targets have several uses outside the DoD. Most obviously, detection of a much wider range of threats would useful in hospital or remote clinic settings. Other applications include detection of biological targets for industrial or personal use (e.g., food safety, pathogen detection, etc.)
KEYWORDS: synthetic biology, phage, austere environmental detection, biological detection, chemical detection