OBJECTIVE: Develop an innovative technology that enables rapid, on-site screening of"suspicious powders"to determine if a biothreat agent is present and if so, its identity. The technology should also enable concurrent documentation of the sample. DESCRIPTION: Since the mailing of Bacillus anthracis spores in 2001, there have been over 35,000 suspicious powder incidents reported in the US. Each incident causes disruption of activities at the scene of the event and costs tens to hundreds of man-hours before the area can be re-opened to the public. The expense of such a shut-down at some locations such as an airport could run into hundreds of thousands of dollars. An overwhelming majority of suspicious powder incidents are hoaxes and the powders identified are often not biological. Currently, first responders use hand-held laminar flow immunoassays1,2 or, much less often, PCR3 for on-the-scene evaluation of suspicious powders to determine biological identification. These tests are costly; equipment to conduct the assays or evaluate the results can cost thousands of dollars, and each assay conducted costs an additional $15-$25 or more2. Recent advances in highly sensitive label-free imaging of bacteria4, lens-free microscopy5, and micro-fluidics6 techniques potentially offer new approaches to rapid screening of suspicious powders. Innovative, cost-effective methods are sought to conduct rapid, on-site screening of suspicious powders by first response personnel. The methods should be within the capabilities of response personnel without requiring advanced levels of expertise or extensive training. PHASE I: As a proof of concept, a method will be developed that is able to rapidly (1-2 min) determine if there are bacteria or bacterial spores present in a 10-milligram sample of suspicious powder that may be diluted by talcum powder or some other type of inorganic material up to a mass ratio of 1000 inorganic to 1 biological. Specific identification of a bacterial biothreat agent or surrogate will be demonstrated within ~30 minutes. PHASE II: An inexpensive (<$100) light-weight reusable hand-held prototype device will be developed that is capable of rapidly detecting and identifying at least 4 bacterial biothreat agents (e.g., Bacillus anthracis, Brucella spp., Francesella tularensis, Yersinia pestis), and at least one biotoxin (e.g., ricin). Consumables shall cost less than $1 per assay. The device will enable documentation of results obtained from each sample analyzed and rapid, simple reach-back capabilities. A system that also has the potential to detect viruses is a plus. PHASE III: Development of a simple pre-processing step may be required to clean-up environmental samples prior to loading into the device developed in Phase II. The resulting system must be usable by first responders and military personnel in full chem/bio protection gear. PHASE III DUAL USE APPLICATIONS: Civilian and military first responders; derivative versions of this technology could be used by point of care clinicians for rapid diagnosis of infectious organisms. REFERENCES: 1. https://www.advnt.org/cart/categories/Biowarfare-Detection/ 2. http://www.advnt.org/products/biowarfare/techlibrary/ComparisonSheet.pdf 3. http://www.idahotech.com/RAZOREX/ 4. Rohit Bhartia, Everett C. Salas, William F. Hug, Ray D. Reid, Arthur L. Lane, Katrina J. Edwards, and Kenneth H. Nealson. 2010. Label-Free Bacterial Imaging with Deep-UV-Laser-Induced Native Fluorescence Appl. Environ. Microbiol. 76(21):72317237. 5. Lee M, Yaglidere O, Ozcan A. 2011. Field-portable reflection and transmission microscopy based on lensless holography. Biomed. Opt. Express, 2(9):2721-2730. 6. Ehrlich DJ, McKenna BK, Evans JG, Belkina AC, Denis GV, Sherr DH, Cheung MC. 2011. Parallel imaging microfluidic cytometer. Methods Cell Biol. 102:49-75.