The Knowledge Foundation, Washington, DC
June 15-16, 2006
Thursday, June 15, 20068:15 Registration, Coffee, Pastries and Exhibit/Poster Setup
8:50 Welcome Address. Opening Remarks
9:00 Common Requirements to Detection Instrumentation - The End User Perspective
Richard L. Dessin, Hazardous Materials Specialist, U.S. Capitol Police
Technology and Level of Training has vastly improved over the last decade and so has the capability of the End User. Provide a better understanding of the needs and the abilities of the End User. Knowledge of the modern End User will help to target technology and device developers and manufacturers working in this industry segment. Detection and Characterization of New and Reemerging Viral Diseases 9:30 Marburg Virus Outbreak in Angola: Operation of a Field Laboratory by the Public Health Agency of Canada - A Case Study
Allen Grolla, Head, Molecular Diagnostics, Special Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada*
In March of 2005, Marburg virus was found to be the cause of an outbreak of hemorrhagic fever that was centered in Uige, Angola. As part of the international assistance team, co-ordinated by the World Health Organization, the Public Health Agency of Canada operated a field laboratory based at the Uige Provincial Hospital from April to June, 2005. The laboratory tested a variety of material from patients, victims and suspicious deaths using SYBR Green I based real-time RT-PCR assays performed on the Smartcycler platform. The lab was able to provide diagnostic test results routinely within 3 hours of sample receipt, which were used to help determine appropriate patient care and funeral procedures and in epidemiological tracing.
*In collaboration with: L.Fernando, U.Ströher, J.Strong, S.Jones, H.Feldmann, NML-PHAC, Canada 10:00 Next Generation Cell Sensors for Virus Identification - Field Based Assays
Spiridon Kintzios, PhD, Associate Professor, EMBIO/AUA, Greece; and Co-Founder, BERADiagnostics Inc.
The state of the art of portable, field-deployable cellular biosensors is presented. An emphasis is given on a new technology called Molecular Identification through Membrane Engineered cells (MIME). The surface of MIME cells has been modified by the artificial insertion of molecules (such as antibodies) that can react in an absolutely specific pattern with potentially any virus under assay. Examples are given regarding the detection of Hepatitis B and C viruses, prions and avian influenza virus using Bioelectric Recognition (BERA) sensors loaded with MIME cells. 10:30 Refreshment Break, Exhibit/Poster Viewing 11:00 Large Scale Genomic Profiling of Influenza
Speaker to be confirmed 11:30 Motif Fingerprint Discovery for H5N1 and Other Lethal Pathogens
Maricel G. Kann, PhD, National Center for Biotechnology Information, NIH
Of the more than 1,500 pathogens known to cause human illnesses, influenza continues to be the leading disease in terms of overall mortality. We developed a genomic survey that quickly differentiates co-circulating influenza strains and identifies specific profiles that allow the recognition of H5N1 avian influenza. We present the epidemiological significance of our analysis and the importance of the motif fingerprint discovery for the surveillance of viral threats.
*In collaboration with: W.Valdivia-Granda, J.Malaga, F.Larson, M.Londono, Orion Integrated Biosciences Inc. 12:00 Metagenomic Analyses of Uncultured Environmental Viral Communities
Mya Breitbart, PhD, Biology Department, San Diego State University
Metagenomic sequencing of uncultured viral communities from seawater, marine sediments, human feces, and human blood show that viral communities are extremely novel and diverse. Through these analyses, we have gained insight into the identity, diversity, and geographical distribution of viruses. Genomics of environmental viral communities allows for the detection of known pathogens and provides the opportunity for identification of emerging threats. 12:30 Luncheon Sponsored by Wohlers & Tan
Marketing Communications for the Sciences Food and Blood Safety: Pathogens and Toxins 2:00 A Novel Method for Detection and Amplification of TSE Diseases for Food and Blood Safety
Alan S. Rudolph, PhD, MBA, Chief Executive Officer, Adlyfe Inc.
We have developed a novel method of detecting and amplifying prion misfolded proteins associated with infectious transmissible spongiform encephelopathies such as BSE (or Mad Cow Disease) and new variant CJD that pose threats to the food and blood supply. Our diagnostic approach is to mimic the protein conformational changes associated with disease by creating synthetic peptide conformational mimetics. These small peptides are conjugated with conformationally sensitive fluorophores that transduce and amplify a signal in the presence of the misfolded protein target. We have demonstrated detection in blood and tissue of both endemic disease and controlled animal models of infection and are developing an ante-mortem blood screening test for detection in animals and humans. 2:30 Detection of Proteinaceous Toxins in Food
Eric A.E. Garber, PhD, Research Chemist, Center for Food Safety & Applied Nutrition, U.S. Food and Drug Administration
The detection of proteinaceous toxins in food is often described as analogous to searching for a needle in a haystack. Methods that have been successfully employed in the detection of proteinaceous toxins can be divided into those which are agent specific and those which simultaneously analyze for multiple toxins at once (multiplex). Agent specific assays employed include ELISAs, a novel 96-well electrochemiluminescence assay, and lateral flow devices (LFDs). The Luminex (BioPlex) multiplex assay has been successfully applied to the simultaneous detection of abrin, botulinum toxin, ricin, and SEB in beverages. Advantages associated with the Luminex multiplex assay include built-in confirmatory tests and the inclusion of internal controls for quantitation and confirmation of assay operation. 3:00 Abscription®- Based Toxin Detection
David McCarthy, PhD, Chief Science Officer, Ribomed Biotechnologies, Inc.
A detection assay for Staphylococcal enterotoxin B (SEB) was developed using the isothermal Abscription® (Abortive Transcription) signal generation system. Abortive transcripts (abscripts®) were produced from an APC (abortive promoter cassette) attached to an SEB specific antibody. The abscription® based assay was more specific than an ELISA assay using the same antibody. The multiplexing properties of abscription® also allow 2-signal signatures to be defined for SEB and other toxins. 3:30 Refreshment Break, Exhibit/Poster Viewing Use of the Host for Sampling and Pathogen Detection 4:00 Universal “Front End” Pre-Assay Sample Purification/Concentration for Nucleic Acid and Immunological Detection Systems
James Meegan, PhD, Senior Director of R&D; and Jeff Rossio, PhD, Program Manager, Invitrogen Federal Systems
Samples containing biothreat agents from clinical and environmental sources often pose two serious problems. First, the agent is usually present in very low concentrations that are below the detection limit of many currently available assays. Second, the sample may contain impurities that interfere with sophisticated modern detection assays. In the case of PCR assays, improved technologies would be particularly desirable to concentrate and purify the sample before nucleic acid extraction. These problems are exacerbated with the utilization of miniaturized bioassay platforms. Typical micro and nano-liter sized samples, in order to contain, for example, a single bacterium, would require a concentration of tens or hundreds of millions of organisms per ml of blood in the starting sample. One way to circumvent these problems is to perform appropriate preprocessing. We will review data on a number of automated concentration/purification technologies to aid with detection of biothreat agents from clinical and environmental samples. Some of these technologies could be envisioned as a universal, “front-end”, pre-assay procedure that will yield a purified and concentrated sample for further analysis in any of a wide array of immunological or nucleic acid/PCR-based detection systems. 4:30 Using “Smart” Holograms as Diagnostic Sensors
Michael J. Powell, PhD, President, Smart Holograms Inc.*
The need for rapid detection of biological and chemical agents is well established. “Smart” holograms are novel sensor systems which use holography as the transduction method for the detection of physical or (bio)chemical stimuli. The sensors are extremely robust, inexpensive and can be used in a number of formats. Sensor holograms will be described for the rapid detection of pathogenic bacteria (e.g. anthrax), antigens, enzymes and a variety of chemical agents.
*In collaboration with: S.Kabilan, A.J.Marshall, A.M.Horgan, C.D.Creasey, Smart Holograms, United Kingdom Equipment Validation and Training Issues 5:00 The Use of Irradiated Bacterial Agents for Validation and Training
Leslee Shuttleworth, Biologist, Emerging Bacterial Diseases, National Microbiology Laboratory, Public Health Agency of Canada
While ramping up their capabilities for biodetection, Canadian agencies involved have identified a common issue related to the inability to validate the equipment purchased and train using actual “A” list agents. The National Microbiology Laboratory in Winnipeg Manitoba chose to address this issue with the use of irradiated bacterial agents. The amount of gamma irradiation was determined for Bacillus anthracis, Yersinia pestis, Francisella tularensis and Brucella sp. The irradiated agents were then tested on various pieces of equipment utilizing different detection methods i.e. RT-PCR and antibodies. Specific training aids have been prepared and utilized for scenario- based training.
5:30 Panel Discussion:
Beyond Detection - Classifying Pathogens and Moving into Therapeutics
6:15 End of Day One Friday, June 16, 2006 8:15 Coffee, Pastries and Exhibit/Poster Viewing Nanotechnology, Genomic and Computational Methods for Pathogen-Specific Identification 9:00 Suspended Microchannel Resonators for Biomolecular Detection
Scott Manalis, PhD, Associate Professor, Dept of Biological and Mechanical Engineering, Massachusetts Institute of Technology
We are developing a new detection method where specific biomolecules adsorb to the walls of a suspended microchannel resonator (SMR) and thereby lower its resonant frequency. Confining the fluid to the inside of the resonator significantly increases sensitivity by eliminating high damping and viscous drag. In this presentation, I will introduce the resonator, show recent progress towards achieving its fundamental limit of detection, and discuss applications for real-time biomolecular detection. 9:30 The NRL cBASS™: A Versatile Biosensor Platform for Orthogonal Assays in Complex Matrices
Lloyd J. Whitman, PhD, Head, Surface Nanoscience & Sensor Technology Section, Naval Research Laboratory
The Naval Research Laboratory’s compact Bead Array Sensor System (cBASS™) is under advanced development for multiplexed detection of pathogens and toxins in environmental and clinical samples. The portable cBASS™ prototype includes a suite of patented and patent-pending technologies for microfluidic, microbead-based microarray assays performed on a Bead ARray Counter (BARC®) sensor chip. Multiplexed hybridization assays and immunoassays can both be performed, and have even been performed simultaneously on the same chip. 10:00 Gene Expression in Vaccinated Volunteers Challenged With Dengue Virus
Vicky J. Gunther, DVD Immunology Section Head, Division of Communicable Diseases & Immunology, Walter Reed Army Institute of Research
Recent advances in molecular biology have led to the ability to measure global changes in gene expression using microarrays; a valuable tool that can help elucidate the mechanism of disease progression. Using the Affymetrix human U133A DNA chips we have evaluated the changes in gene expression that occur in vaccinated human volunteers and unvaccinated controls after challenge with either dengue 1 virus (DENV-1), or dengue 3 virus (DENV-3).
*In collaboration with: M.Vahey, R.Das, R.Hammamieh, M.Jett, R.J.Putnak, S.Fernandez, D.R.Palmer, A.G.Lyons, K.H.Eckels, S.J.Thomas, R.V.Gibbons, and W.Sun, WRAIR; and D.W.Vaughn, US Army Medical Research & Materiel Command 10:30 Refreshment Break, Exhibit/Poster Viewing 11:00 Integrated Detection of Viral Pathogens and Early Infection Biomarkers
Willy A. Valdivia-Granda, PhD, CEO and Founder, Orion Integrated Biosciences Inc.*
We developed an integrated computational analysis system (ICAS) for the identification and classification of viral pathogens. Hybridization signals produced by our liquid barcoded nanoarrays can classify accurately hundreds of viral species including Poxviruses, Flaviviruses, Alphaviruses and Influenza. ICAS was also used to recognize day-specific and infection-dependent transcriptional interactions formed from gene expression profiles of cynomolgus macaques challenged with variola virus. Our analysis represent a paradigm shift in the detection and characterization of known and unknown pathogens and the identification of early infection biomarkers in human patients exposed to smallpox and other biological threats.
*In collaboration with: J.Malaga, F.Larson, M.Londono, Orion Integrated Biosciences Inc. 11:30 Canary B-Cell Sensor for Rapid, Sensitive Identification of Pathogens
James D. Harper, PhD, MIT Lincoln Laboratory
We have harnessed nature’s pathogen identifier, the B lymphocyte, to enable detection of 50 cfu of pathogens (including anthrax spores) in less than 3 minutes. Tests using this cell-based sensor technology, which we call CANARY, are very simple to perform in inexpensive equipment and provide a unique combination of extreme speed and sensitivity that can dramatically enhance pathogen identification capabilities for bioaerosol monitoring, medical diagnostics, and other applications. 12:00 Broadband Detection of Biological Agents Using Single-Molecule Genomic DNA Mapping
Jenny E. Rooke, PhD, Vice President, U.S. Genomics, Inc.
Conventional approaches to biological agent detection employ threat-specific reagents which can be defeated by naturally arising and/or engineered variation. U.S. Genomics’ novel biosensor technology uses a single reagent set to detect a range of biological agents. The Company’s Direct Linear Analysis (DLA) technology performs high-throughput mapping of genomic DNA to reveal patterns unique to each organism. DLA is robust against variations that foil conventional assays, enabling detection of novel and engineered biological threats. 12:30 Lunch on Your Own Physical Methods for Pathogen Identification and Imaging 2:00 Multiplexed Electrochemiluminescence Detection of Biowarfare Agents
George Sigal, PhD, Director of Chemistry, Meso Scale Defense
Meso Scale Diagnostics, LLC. (MSD) develops and manufactures a line of ECL-based detection systems. MSD’s systems can perform, in one well, multiplexed measurement for up to 100 analytes of interest. MSD has already demonstrated the use of this technology for ultra-sensitive multiplexed immunoassays for BWAs (bacteria, viruses and toxins). MSD has developed a product line that extends this detection technology from the front line/ first responder all the way to the central laboratory setting with equivalent performance across all instruments. 2:30 Rapid, Multiplexed Biodetection using Luminex® xMAP™ Technology
Sherry A. Dunbar, PhD, Senior Director, Biology R&D, Luminex Corporation
The Luminex® xMAP™ system couples bioassays with advanced digital signal processing and proprietary identification techniques to perform multi-analyte testing of up to 100 features in real time. The general configuration of an xMAP™ assay can be described as a suspension array where specific capture moieties are covalently coupled to the surfaces of internally dyed microspheres. Fluorescent dyes contained within the microspheres provide unique spectral characteristics allowing each microsphere set to be distinguished from all others in the multiplex. The target is labeled for fluorescent detection. Examples will be presented describing rapid, multiplexed protein and nucleic acid analyses for infectious agents that demonstrate the utility of the platform. Benefits of the system include speed, economy, flexibility, and advanced capabilities. The potential for simultaneous detection of tens, hundreds, and even thousands of protein and nucleic acid targets provides for simultaneous, rapid, sensitive, and specific molecular analyses. 3:00 Development of an Integrated Detection and Identification System for Airborne Biological Agents
Mary Beth Tabacco, PhD, Vice President, Director of Research; and
Jay Lewington, PhD, Director, Biological Detection Technology, Smiths Detection
Smiths Detection is developing sensors and integrated instruments for rapid detection and identification of airborne biological agents. A biosensor array has been developed that provides real-time detection and classification of microorganisms based on molecular recognition and fluorescence spectroscopy. This biosensor is being integrated with an identification instrument that is based on a novel approach for surface plasmon resonance and light scattering. This device can continuously monitor for up to 20 biological agents and provide identification in 15 minutes or less. 3:30 Refreshment Break, Exhibit/Poster Viewing 4:00 Deep UV LED Lamps for Biodetection
Yuri Bilenko, PhD, Optoelectronic Products Manager, Sensor Electronic Technology, Inc.*
Light Emitting Diodes (LEDs) have numerous advantages for applications as light sources for biosensors at wavelengths below 365 nm due to their fast switching times, low noise, and excellent capabilities for meeting system design requirements. The LEDs allow for the spatial and time integration of multiple wavelengths in one sensor, with embedded intensity and spectral feedback. We report on deep UV LEDs (UVTOP®) developed and manufactured at Sensor Electronic Technology, Inc. (SET) that have been successfully tested in a number of biodetection and bioanalytical systems.
*In collaboration with: J.P.Zhang, X.Hu, A.Lunev, J.Deng, T.Katona, M.S.Shur, and R.Gaska, Sensor Electronic Technology 4:30 AFM Measurements to Distinguish Bacillus Anthracis Spores from Those of Related Species
Larry W. Burggraf, PhD, Professor of Chemical Physics and Engineering Physics, Air Force Institute of Technology
Bacillus spore surface morphology was imaged using atomic force microscopy (AFM) to distinguish between four closely related species; Bacillus anthracis Sterne strain (BA), Bacillus thuringiensis var. kurstaki (BT), Bacillus cereus strain 569, and Bacillus globigii var. niger. Phase images of spore surface morphology are rich with information. Force distribution measurements for BA-aptamer interactions with BA spore surfaces and BT spore surfaces demonstrate potential for using modified AFM probes to distinguish surfaces of different spore species. 5:00 Label-Free Colorimetric Detection of SNP in PCR Products
Huixiang Li, PhD, Research Associate, Dept of Chemistry, University of Rochester
Single-stranded DNA adsorbs on negatively charged gold nanoparticles (Au-nps) with a rate that depends on sequence length and temperature. After ss-DNA adsorbs on Au-nps, we find that the particles are stabilized against salt-induced aggregation. These observations can be rationalized on the basis of electrostatics and form the basis for a colorimetric assay to identify specific sequences and single nucleotide polymorphisms on polymerase chain reaction (PCR)-amplified DNA. The assay is label-free, requires no covalent modification of the DNA or Au-np surfaces, and takes on the sensitivity of PCR. Most important, binding of target and probe takes place in solution where hybridization occurs in less than 1 min. 5:30 Selected Oral Poster Presentations
6:00 Concluding Remarks. End of Conference
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