BIODETECTION TECHNOLOGIES: Identification Innovations and StrategiesMay 1 - 3, 2002 • Hilton Alexandria Mark Center • Alexandria, VA USA [ Overview | Agenda | Call for Posters | Register ] View Electronic Brochure (PDF)OverviewQuick and accurate assessments of biological agents has become a focal point within the biosensing community. This conference will explore the latest research and state-of-the-art technologies of biological detection of these agents. As the demand for these identification systems increases, it is vital for you to gain access the latest information available. Our select group of international experts will specifically address such topic areas as:Biosensor Technology * Taste Chips Bioanalysis Technology * Cytometery Rapid Detection * PCR-Based Detection Instruments and Mark your calendars not to miss this networking opportunity to learn of the latest in biological detection. Register today!RELATED LINKS AgendaWednesday, May 1, 2002
Preconference WorkshopIntegrating Biological and Chemical Detection Technology: Today's Solutions and Tomorrow's Issues1:00 Registration/Refreshments 1:50 Opening Remarks 2:00 Optical Detection Capabilities for Biological and Chemical Agent Aerosols This presentation will provide an overview of a new-start effort to develop detection capabilities for biological and chemical agent aerosols. This project is a cooperative effort involving R&D groups at the Edgewood Chemical and Biological Center, Aberdeen, MD, MIT-Lincoln Laboratory, Lexington, MA, and the Naval Research Laboratory, Washington, DC; and is funded through CB Tech-Base (US Army). The project goal is to demonstrate improved rapid biological aerosol discrimination and rapid chemical agent particle detection in a single device using a combination of primarily optical technologies. 2:30 Chemical Biological Mass Spectrometer (CBMS) Mass spectrometers currently play a major role in the detection of chemical and biological agents on the battlefield. Two items, the MM1 mobile mass spectrometer and the Block I Chemical Biological Mass Spectrometer (CBMS) are representative devices currently in the field. More recently, the Soldier and Biological Chemical command has developed a Block II CBMS for the detection and identification of warfare agents in reconnaissance and point detection roles. This rugged, user-friendly detector has enhanced performance bands and combines many of the features found in the earlier instruments. 3:00 The Block II Chemical Biological Mass Spectrometer (CBMS): Issues and Solutions in Integrated Chemical-Biological Agent Detection Integrated detection and identification for both chemical and biological warfare agents imposes additional requirements above those for doing either class of agents alone. Among the issues are the specific sampling systems and their interface with the detector, sample pretreatment process, the modes of operation of the detector, and the means of agent identification. This presentation supplements the introductory CBMS presentation in describing how these requirements are met in the Block II CBMS. *In collaboration with M. B. Wise, K. J. Hart, S. A. Lammert, C. V. Thompson, D. A. Wolf, M. N. Burnett, A. A. Vass, I. B. Robbins, and D. A. Clayton, ORNL. 3:30 Refreshment Break 4:00 Detecting Biological Agents in Mail Using Fluorescence Particle Sizing* Fluorescence particle sizing has been demonstrated over the last few years to be the most effective way to detect airborne biological particles in near real time. It is based on the ability to count and size airborne respirable particles combined with the ability to distinguish biological particles based on the fluorescence emission from specific biomolecules. Most tests of equipment have occurred outdoor where the background is fairly benign. However, detecting biological releases from mail in the postal facilities presents a daunting task because of high background total and fluorescent particle counts combined with intermittent operation of the sorting equipment and the localized nature of the releases from the mail. This presentation will discuss how we have succeeded in modifying our fluorescent particle sizing equipment to detect low emissions of biological simulants from mail in sorting facilities while avoiding false alarms from the intermittent operation of the equipment. *In collaboration with Steven Herz, Computing Devices Canada. 4:30 BioHaz-Field Screening for Biological Agents Attendees will be introduced to and provided an opportunity to experience hands-on techniques for a new and inexpensive biological detection system. The BioHAZª is a portable field system designed for use by emergency response personnel at incidents where biological materials may be present and sampling collection and analysis is necessary. The system contains all materials necessary to collect air, liquid and solid samples for on-site biological analysis, and prescribes detailed instructions to ensure sample integrity, resulting in legally defensible data. The system is designed to allow the user to select the proper sampling package which includes sample sponges, sterile swabs, syringes, spatulas, bags, and culture tubes; chain-of-custody forms, markers, and tamper seals. Analysis is conducted through fluorometry, luminesence, colorimetry, and sensitive membrane antigen rapid tests. Step-by-step instructions from preparation for entry into the hot zone through sample splits, analysis and transfer to law enforcement are included. The system is packaged in rugged portable watertight containers with each sample collection kit and sample-processing kit marked for easy locating and use. All sampling/processing kits and other individual components are designed to be disposable and replaceable. 5:00 Panel Discussion on Issues and Solutions on Combining Biological and Chemical Technologies 5:30 End of Workshop Main ConferenceThursday May 2, 20027:30 Registration, Poster/Exhibit Set-Up, Coffee and Pastries 8:25 Opening Remarks 8:30 Key Note Address - Department of Defense Biodetection Research, Development and Acquisition: Current and Emerging Technologies for Detection of Biological Agents for the Warfighter for Military Operations and Homeland Security Department of Defense seeks to ensure U.S. military personnel are equipped to operate in future battle spaces that may feature biological contamination. Recent events have shown that the concept of future battle spaces may be changing. This presentation will describe current DoD biodetection technologies to support the warfighter in a variety of environments, research & development to address current technological limitations, and some of the challenges and opportunities in supporting civilians as part of homeland security. Biological Detection I9:10 Chairperson's Remarks 9:15 Strategies for the Detection of Unknown Biological Materials Current strategies for the detection and identification of biological agents depend on known biological properties - culture characteristics, specific biochemistries, specific antigens, and specific nucleic acid sequences. What strategies can be implemented for detection of unknown agents? Detection and identification strategies, as they might be applied to the detection of unknown materials, will be reviewed as to speed, complexity, and information generated. Trade-offs among these parameters and the introduction of new detection and identification schemes in concert with current, proposed, or future technologies will be discussed. The intent of this paper is not to solve the problems, but to provoke new ideas, so that effective biological detection capabilities for the 21st century can be developed. Paper provided by Peter J. Stopa, Ph.D., Director, Marine Corps Chimical/Biological Technology Base, US Army Edgewood Chemical Biological Center. 9:45 Testing for Anthrax at the Armed Forces Institute of Pathology Abstract not available at time of printing. 10:15 Refreshment Break and Poster/Exhibit Viewing 10:45 Biodetection With Flow Cytometry: Better, Faster, Cheaper The objective of our research is to test the feasibility of an innovative approach to rare cell detection using a CCD prism spectrometer to image an entrained flow through a rectangular glass tube. This approach is especially useful for the detection of rare cells where a high volumetric flow rate is desired. We present the results of our work with the detection of Escherichia coli O157:H7 carrying multiple fluorescent labels. This technique has key advantages over current alternatives, including: (1) high volumetric flow rate, (2) capability of detecting single microorganisms (3) automatic operation, and (4) easy implementation in a rugged, portable system. This project was funded by NSF STTR grant DMI-9810567 (Phase I and Phase II), NASA Space Grant NGT 40050, and NSF REU grant 9732039. * In collaboration with Amanda S. Votaw, and Tony Deromedi, Department of Physics and Astronomy, University of Wyoming. 11:15 Nucleic Acid-Based Detection of Antibiotic Resistant Biowarfare Agents Using the Roche Lightcycler The use of ciprofloxacin and tetracycline (doxycycline) to prevent and treat infectious disease caused by agents of biowarfare has raised concerns over early detection of antibiotic resistant organisms. Ciprofloxacin resistance (Cip) is generally due to the mutation of one or more cellular topoisomerases. In contrast, tetracycline resistance (Tet) is usually mediated through the acquisition of new genetic material. We have developed individual 5'nuclease assays that can identify Tet gene classes A, B, C and D as well as a single probe-set that can identify Cip Yersinia pestis. The sensitivity of our Tet and Cip assay is on the order of 1-10 CFU of bacteria. Taken together, our results indicate that the use of fluorescent probes in DNA detection assays can offer significantly increased speed, sensitivity and classification of antibiotic resistant phenotypes in agents of biowarfare. *In collaboration with Wei Fan, Walter Reed Army Institute of Research. 11:45 Lunch Sponsored by MesoSytems Technology 1:10 Fighting Germs on the Front lines; An Integrated Laboratory Approach to Field and Lab Analysis and Surveillance Proactively testing for different infectious agents can provide valuable data and insight to commanders and health officials. Through use of advancements in information technology, rapid diagnostics can be used as surveillance tools. The L.E.A.D.E.R. (Lightweight Epidemiology and Advanced Detection, and Emergency Response) System is a medical surveillance tool providing real-time analysis of medical data linked to real-time and accurate laboratory data. L.E.A.D.E.R. system is a suite of tools that provides timely diagnostics and consultative capability in a light, highly mobile platform. 1:40 Real-Time Detection of Biological Aerosols by the Matrix-Assisted Laser Desorption/Ionization and On-Line Aerosol Time-of-Flight Mass Spectrometry One of the recognized problems in current protection against biological warfare agents is the lack of a bioaerosol alarm. At TNO we are developing a system based on particle preselection and matrix-assisted laser desorption/ionisation (MALDI). Using this combination, mass spectra were obtained from single biological aerosol particles using an aerosol time-of-flight mass spectrometer (ATOFMS). The inlet to the ATOFMS is coupled with an evaporation/ condensation flow cell that allows the aerosol to be coated with matrix material as the sampled stream enters the spectrometer. Mass spectra were generated from aerosol composed of gramicidin-S or erythromycin, two small biological molecules, or from aerosolised spores of Bacillus subtilis var niger. Three different matrices were used: 3-nitrobenzylalcohol, picolinic acid and sinapinic acid. A spectrum of gramicidin-S was generated from approximately 250 attomoles of material using a molar ratio of 3-nitrobenzylalcohol to analyte of approximately 20:1. A single peak, located at 1224 Da, was obtained from the bacterial spores. The washing liquid and extract solution from the spores were analysed using electrospray mass spectrometry and subsequent MS/MS product ion experiments. This independent analysis suggests that the measured species represents part of the B. subtilis peptidoglycan. The on-line addition of matrix allows quasi-real-time chemical analysis of individual, aerodynamically sized particles, with an overall system residence time of less than 5 seconds. These results suggest that MALDI ATOFMS can provide nearly real time identification of biological aerosols.*In collaboration with B.L.M. van Baar, Ch. E. Kientz, TNO Prins Maurits Laboratory, The Netherlands and M.A. Stowers, A.L. van Wuijckhuijse and J.C.M. Marijnissen, Delft University of Technology, The Netherlands Biosensors2:10 Chairperson's Opening Remarks 2:15 Solution Based Analysis of Multiple Analytes by a Sensor Array: Towards the Development of an "Electronic Taste Chip" A new type of sensor array that functions as an "electronic taste chip" has been designed fabricated and tested. The hybrid micro-machined structures have been interfaced directly to charged-coupled-devices, which are used for the simultaneous acquisition of colormetric data from the various "taste bud" elements. This universal platform has been applied successfully to the analysis of complex fluids containing acids, bases, solvents, antibodies, antigens and various DNA oligomers. 2:45 Real Time/Near Real Time Biosensor Detection of Bioterrorism Agents Conventional procedures to detect pathogenic microorganisms in food, water, and the environment can take days to perform and may provide inconclusive identification. Fiber optic biosensors can identify specific pathogens and toxins in minutes, instead of days, directly from complex matrices using robust antibody-, receptor-, and/or nucleic acid-based assays. Furthermore, biosensor assays can be simplified and automated for field use with a portable unit by minimally trained personnel, and can be used for real time/near real time detection of bioterrorism agents. 3:15 Refreshment Break and Poster/Exhibit Viewing 3:45 Development of Biological Sensor Systems Based on Array Immunosensors Prototype sensors tend to employ components with refined capabilities that are generally costly and tend to exceed the requirements of a commercialized field system. Moreover, prototype systems usually require drastic reconfiguration in order to accommodate the needs of the end user, for example portability, easy sample introduction, simplified data processing and read out of results. In this presentation, we will describe the development of array immunosensors with defense applications from laboratory prototypes to commercial products. *In collaboration with Timothy Postlethwaite, Ron Priest, Peng Zhang, and Rich Sorrells, Constellation Technology Corp. 4:15 Semi-Selective Optical Sensors for Real-Time Detection of Biological Warfare Agents Echo Technologies is developing a suite of optical sensors to detect and distinguish broad classes of microorganisms including bacteria, spores, toxins and viruses. This approach is a departure from other developmental sensors designed to identify a single organism or chemical, and is therefore referred to as semi-selective. This approach is particularly valuable when faced with an unknown or genetically modified organism. The sensors are based on the use of molecular recognition and fluorescence spectroscopy. Detection has been demonstrated for both liquid and aerosolized samples. A handheld BW detection system is presently being tested in the laboratory. 4:45 Sensor Data Integration and Management The marketplace is full of a variety of best of breed sensors: triggers, collectors, detectors, identifiers, alarms, and the like. In and of themselves, they all perform very specific jobs for their defined threat and environment. What we see missing is aggregation of sensor data into a complete situational awareness picture. What we have is a solution that is sensor-agnostic a standard platform (much like Windows/Intel) that can be used to plug and play sensors (much like computer PC peripherals) independent of environment and threat and which also provides a facility to integrate the disparate data that is generated from potentially very disparate (in type and geography) sensor networks. 5:15 End of Day One Friday May 3, 20028:00 Poster/Exhibit Viewing, Coffee and Pastries 8:55 Opening Remarks 9:00 Special Presentation - Tapping Into a $550 Million Technology R&D Funding Opportunity: A Primer on the Department of Defense SBIR Program We will present a brief overview of the Department of Defense (DoD) Small Business Innovation Research (SBIR) program and focus on latest developments in the program. DoD awards over $550 million in R&D contracts to small technology companies each year. Many improvements in the program make it an attractive source of early-stage capital for small companies interested in developing commercially viable technologies. Any small business entrepreneur, potential investor, or commercial strategic partner involved in high-technology R&D is likely to be keenly interested in learning more about the SBIR program. The Small Business Technology Transfer (STTR) program-which funds cooperative R&D projects between small business and non-profit research institutions will also be briefly covered. Rapid Detection Technologies9:40 Chairperson's Opening Remarks 9:45 Rapid PCR-Based Detection Instruments and Chemistries PCR is particularly useful for detecting microorganisms. Fluorescence-based quantitative PCR is now available and there are several thermal cycling devices designed for real-time PCR monitoring. However, current fluorescent reporting technologies have limited speed and sample pre-processing is slow. Improvements to PCR for application in field-based detection will be discussed. These will include: operator-friendly sample processing; fast cycling using electrical conducting polymer; and novel fluorescent detection methods called "ResonSense" and "Angler". 10:15 Rapid Sample Processing Integrated with Real-Time PCR Utilizing meso-and microfluidic functionalities and a novel random access rotary valve, the GeneXpert cartridge system fully integrates all sample-processing steps with real-time PCR. After the specimen is added to the disposable cartridge, it is sealed and placed in a processor. Results are available in 30 minutes or less. Up to 6 mL of specimen can be processed in the first generation cartridge, and clinical and environmental applications have been demonstrated for Group B Streptococcus, anthrax, and tuberculosis. PCR product can be accessed for post-PCR analyses. 10:45 Refreshment Break and Poster/Exhibit Viewing 11:15 US Army Science and Technology Objective to Develop Tests for Rapidly Detecting Microbial and Chemical Contaminants in Food and Water Army medical personnel and US forces in the field require devices and equipment to rapidly detect bacterial and chemical contaminants in food and water sources to validate their safety for consumption. Current field-testing is very limited and may take days to complete. Samples sent to reference laboratories may take weeks for testing, during which time consumption of the commodities may be discouraged or prohibited. This Science and Technology Objective (STO) focuses on exploiting and applying new technologies in chemistry, genetics and immunology to create new field medical products that will rapidly detect and identify contaminants in water. This includes bacteria and other microbes associated with fecal contamination, agricultural pesticides, and toxic industrial chemicals. To achieve its goals, this Army STO is leveraging with multiple partners, such as Department of Defense programs, academia, industry, federal laboratories, and regulatory agencies. *In collaboration with Paul L. Knechtges and W. D. Burrows, U.S. Army Center for Environmental Health Research. 11:45 Rapid Assays for Biothreat Agent Detection for Public Health Labs In the event of an attack using biological weapons, there is a need for rapid detection and identification of bioterrorism agents. This is the mission of the Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory of the Bioterrorism Preparedness and Response Program, CDC. For nucleic acid detection and identification of bioterrorism agents, the BRRAT laboratory currently uses PCR-based 5' fluorogenic nuclease assay detection run on a variety of instrument platforms including the ABI7700, Roche Light Cycler and Cepheid Smart Cycler. Antigen detection assays using time-resolved fluorescence are performed on the Wallac Victor2 instrument. The list of possible bioterrorism threat agents continues to grow and rapid nucleic acid and antigen detection assays are in development for many agents. 12:15 Lunch on Your Own 1:40 Rapid On-Site Biological Sampling and Detection for First Responders The United States has experienced an onset of hundreds of biological hoaxes since 1997 and actual Anthrax attacks in 2001, initially resulting in overreactions of emergency response agencies to hoaxes and spending in excess of $10 M. Current Anthrax attacks through the postal service are totally overwhelming local, state and federal HAZMAT teams attempting to respond to all threats as significant credible threats. A rapid, simple and inexpensive solution was requested by these agencies in 1999 to help solve this still occurring problem. The immediate solution to this problem is the BioHAZª, a portable field system designed for use by emergency response personnel at terrorist incidents where biological materials may be present, and sample collection, screening and possible identification is warranted. This presently utilized system contains all materials necessary to collect liquid and solid samples for on-site biological screening and off-site positive analysis, and prescribes detailed instructions to ensure sample integrity, resulting in legally defensible data. 2:10 The Utilization of Detection Systems by First Responders to Screen Field Samples for Suspected Biological Agents BW agent rapid detection or identification systems have primarily focused on testing defined aerosol samples released under favorable, controlled conditions or controlled samples assayed under laboratory conditions. Although these methods may provide information on the sensitivity, cross-reactivity, and some interference factors, they may not be practical for use by Fire and Police HazMat teams under field conditions. The events following 9/11 have placed local responders in situations where hoax material or "false alarms" are the norm. In these instances many hundreds of citizens await word from local officials whether they must be field decontaminated, begin antibiotic treatment, or are free to go home. Under these conditions, a test must be rapid, easy to perform and be able to rule out false alarm material. The latter point may be the most critical for only 1 or 2 out of several hundred responses may result in an actual positive. Testing with two different technologies previously utilized by various agencies in the field (Hand Held Assays and Luminescence) have shown potential for meeting these requirements when evaluated with common environmental samples as well as when tested with adulterants or interfering substances which may be used by a terrorist. *In collaboration with L. Loomis, J. Tartal, C. Trudil, New Horizons. Biological Detection II2:40 Chairperson's Opening Remarks 2:45 DNA Analysis for Forensic Identification of Strains in Anthrax, Plague and Tularemia Bacterial bioweapons can be identified and tracked through differences in their genomes. The most rapidly mutating DNA regions provide the greatest resolution, even toward differentiation of very closely related isolates. My lab has developed PCR-based DNA fingerprinting analysis for anthrax, plague, tularemia, brucellosis and melioidosis pathogens. In addition, we have been developing large standardized databases containing the genetic identity of strains from worldwide collections. These resources are more than an academic exercise and have been successfully applied to recent bioterrorism events. 3:15 Refreshment Break and Poster/Exhibit Viewing 3:45 Catching the Light: Fast Detection of Low Numbers of Bacterial Cells Using Enhanced Bioluminescence Bioluminescent measurement of ATP provides a simple means for detecting microbial contamination. An amplified endpoint assay based on adenylate kinase (AK) can allow sensitivity to be improved 100-fold and, by using immunocapture to selectively extract and concentrate target cells, specific detection of low concentrations of bacteria can be achieved. The assays have been automated, take 10-15 minutes to complete and provide limits of detection of around 102 cells ml-1. 4:15 Detection of Pathogenic Bacteria by Volatile Organic Compound (VOC) Analysis Developments in rapid detection technologies have made countless improvements over the years. However, because of the limited sample that these technologies can process in a single run, the chance of capturing and identifying a small amount of pathogens is difficult. The problem is further magnified by the natural random distribution of pathogens in foods. Methods to simplify pathogenic detection through the identification of bacteria specific VOC were studied. E. coli O157:H7 and Salmonella typhimurium were grown on selected agar medium to model protein, and carbohydrate based foods. Pathogenic and common spoilage bacteria (Pseudomonas and Morexella) were screened for unique VOC production. Bacteria were grown on agar slants in closed vials. Headspace sampling was performed at intervals up to 24 hours using Solid Phase Micro-Extraction (SPME) techniques followed by GC/MS analysis. Development of unique volatiles was followed to establish sensitivity of detection. E. coli produced VOC that were not found in either Trypticase Soy Yeast agar (TSYA) blanks or spoilage organism samples were - indole, 1-decanol, and 2-nonanone. Salmonella specific VOC from TSYA were 3-methyl-1-butanol, dimethyl sulfide, 2-undecanol, 2-pentadecanol and 1-octanol. Trials on potato dextrose agar (PDA) slants indicated VOC specific for E. coli and Salmonella when compared to PDA blanks and Pseudomonas samples. However, these VOC peaks were similar for both pathogens. Morexella did not grow on PDA slants. *In collaboration with Josh Magnone, Walter Yeomans, Ken Racicot and Edmund M. Powers, U.S. Army Soldier Biological and Chemical Command, Natick Soldier Center 4:45 The Use of Supersensitive P-Chip/Multi Photon Detection (MPD) for Detection of Biological MPD technology is applicable to the full spectrum of BW agents, including viruses, bacteria, algae and biotoxins. Initially, we propose the use of MPD enhanced detection methods for individual targets. We expect to complete the development of supersensitive P-chip/MPD for BW agents within a year. This universal system, able to detect all major groups of biological warfare agents, represents the most sensitive and pragmatic solution to the detection of BW threats. The proposed applications of MPD for the detection of biological warfare agents can be divided into two synergistic projects: use of a panel of MPD enhanced immunoassays for detection of biotoxins, and development of a universal biological warfare agent detector using supersensitive P-chips/MPD. Call for PostersCall for Posters Register
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