REBS The Resource Effective Bio-Identification System

Transcription

1 REBS The Resource Effective Bio-Identification System A.P. Bartko, Ph.D June 25, 2013

2 Effective Biological Identification begins with REBS REBS Attributes Collects and Identifies in minutes without the need for a trigger Effective for bacteria, viruses, and toxins Demonstrated < 10 6 CFU Min/M 3 sensitivity Demonstrated zero False Alarm Rates in actual environmental testing Agility to add new threats in 24 hours Continuous collection Environmental particulate history stored on collection media Non-destructive, sample compatible with genetic confirmation No man in the loop, portable, battery powered, and network ready Hands-free maintenance for up to a month < 1 cent per sample, < $100k per system 2

3 Raman Spectroscopy Incident laser energy is shifted by the molecular bond resonant vibrational energy. Light/matter interaction results in a Raman shifted photon specific to the molecular bond Molecule structural conformation influences spectra and are used for biological identification Biological materials yield complex Raman spectra Application of Raman in REBS DNA/RNA, spore coat, capsule, cell membrane, intracellular contents all contribute to the measured spectra Bonds and conformation both contribute to identification Spectral measurement does not degrade viability/activity Cell viability, in some instances, is discernible in spectra Single cell/particle measurements removes interferant confusion Capability to identify mixed threats Factors influencing growth (extrinsic variability) are discarded No primers, probes, liquids reagents or substrates needed Accurate, agile and economical point identification system 3

4 Biological Spectral Signatures Comprised from Chemical Moieties Vegetative Bacillus subtilis spectral signature shows many Raman shifted peaks that can be attributed to different types of molecular bond resonances Each molecular bond resonance experiences a local environment due to the biological conformational structure. Conformation influences the natural resonance peak position, peak width and peak amplitudes. 4

5 REBS System Description 5

6 REBS Process Animation 6

7 REBS Digital Threat Identification Process Triaged approach to Identification Step 1Organic/Inorganic ( 0.1 sec) Step 2 Bio/Non Step 3 ID (10 sec) ( 50 sec) 7

8 Gov t Testing Summary Location (Sponsor) Date Type Result Fort Bliss, Texas (JPM- G) Dugway, Utah Piggyback (JPM-BD) Battelle Columbus, Ohio (JPM-BD) Battelle Columbus, Ohio (JPM-CA) JHU-APL/ ECBC (JPM- BD) DPG (JPM-CA for CBDS) Boston, MA Subway Test (DHS S&T) West Jefferson,OH, CRADA w/ JPM-NBC 6/2009 to 7/ /2009 to 11/ /2009 to 01/2010 Background sampling TRE Simulant challenge in realistic environment JBTDS Identifier TRE Samples, ID performance with killed BWA Zero False Positives ID sensitivity - 25 ACPLA 100% Identification 6/2009 CWA (particle) ID feasibility Dual threat feasibility proven 9/2010 JBTDS TRE shoot off with simulants 2/2011 to 7/ /2012 to present 06/2013 to 07/2013 ASEC and ABT testing with irradiated materials, unknown biological, near-neighbors and interferants Simulant release in actual environment Live agent integrated system testing and ABT Selected for JBTDS Competitive Prototyping Demonstrated Sensitivity, Specificity, and False Alarm Performance Zero False Positives, 100% identification of simulant release TBD 8

9 Identification Performance Summary BWA agent and simulants used for aerosol Limits of Identification performance determination Bacillus anthracis-ames, Sterne Escherichia coli Bacillus globigii Francisella tularensis Bacillus thuringiensis-kurstaki and ATCC 9727 Botulinum toxoid Bacillus cereus-atcc Ovalbumin grade III and V Brucella suis Bovine serum albumin Burkholderia mallei MS2 bacteriophage Burkholderia pseudomallei T3 bacteriophage Yersinia pestis- Kim, Nairobi, CO92 Venezuelan Equine Encephalitis-Trinidad Erwinia herbicola Vaccinia 2009 Detector TRE Results > 98% Probability of Identification Average system response function (line) 80% confidence interval (gray shaded region) (+) symbol = positive correct ID found within 30 minute interval (o) symbol = no ID found within 30 minute interval Data representative of all threat classes < 25 PPL sensitivity demonstrated 9

10 Actual Environmental Testing Performance Summary Location Fort Bliss, TX Boston, MA Subway Duration 06/ / /2012 4/2013 Hours 740 2,454 Samples 651 5,860 Particles 1,494,557 3,498,363 CH ,488 Bio Simulant + N/A 56 Threat+ 0 0 Limit of Identification < 10 6 CFU Min/M 3, zero false positives Threat Database: Bacillus anthracis, Francisella tularensis, Venezuelan equine encephalitis, Yersinia Pestis, Vaccinia, Bot A and Bacillus globigii (BG) as a BWA simulant 10

11 Challenges Moving Forward REBS is currently an integrated point detection system at a TRL 6 level. Additional operational test and evaluation is needed to demonstrate improvements meet or exceed reliability. Field trials of REBS demonstrated zero false positives with a 95% upper confidence bound of FPR. Further operational testing is needed to demonstrate lower FPR. Test and evaluation methodologies presume the use of liquid media for identifier compatibility with collectors. Testing gaps exist for emerging and reagent-free technologies. Process integration with orthogonal technologies such as Genomic-based identifiers for confirmation. 11