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Detection Technologies Conference DocumentationPublished by: Knowledge Foundation Published: Jan. 1, 2003 - 150 Pages Table of ContentsPre-Conference Workshop Case Studies in Detecting and Analyzing New Threats Monday, December 8, 2003 8:00 Registration, Poster/Exhibit Setup, Coffee and Pastries 8:45 Chairperson’s Opening Remarks Johnny Callahan, MS, MT(ASCP), Scientist - Viral Assay Development, Tetracore, Inc. 9:00 Severe Acute Respiratory Syndrome (SARS) Coronavirus - Molecular Epidemiology and the Diagnosis of the Virus Frederick C. Leung, Dean of Science, Professor, The University of Hong Kong, China Severe Acute Respiratory Syndrome (SARS) is a newly emerged disease caused by a novel coronavirus (SARS CoV) which spread globally in a very short time affecting over 30 countries. We have characterized the SARS CoV complete genome sequences. Based on the SARS CoV-S1 gene, SARS patients’ samples in Hong Kong and Guangdong during February-April 2003 were genetically sequenced and phylogenetically analyzed in comparison with other sequences available from public databases. The majority of the Hong Kong viruses phylogenetically cluster closely together with a single index case started in late February and viruses identified in others Guangdong and Beijing is genetically more diverse. The return of SARS this winter has been warned by WHO and CDC. Therefore, a rapid, reliable diagnosis assay is needed for monitoring the spread of the disease. We developed a method to eliminate false negative results with enhanced sensitivity, based on use of the nucleocapsid (N) gene as target in a one step RT-PCR multiplex real time quantitative PCR assay. In conclusion, nucleocapsid gene represents an additional sensitive molecular marker as compared with the Ib gene target for the diagnosis of the SARS-CoV and can be further adapted into a high throughput platform assay. 9:40 Detecting Threat Agents in Environmental and Clinical Samples: Challenges and Applications James Higgins, Microbiologist, USDA-ARS During the anthrax bioterror event in the Washington, DC area in October, 2001, the USDA-ARS established a field laboratory for testing of swab, air, and mail samples for the presence of spores. Applying molecular biology-based detection regimens to these samples revealed a number of challenges and obstacles that become apparent in “real world” situations. Subsequently, we have been conducting research on ways to improve the sensitivity and specificity of threat agent detection in environmental and clinical samples. 10:20 Refreshment Break and Poster/Exhibit Viewing 10:50 SARS Case in Canada: Virus Isolation, Rapid Diagnostics, Tool Development and Lessons Learned J. Neil Simonsen, BSc, MD, DTM&H FRCPC, Canadian Science Centre for Human and Animal Health, National Microbiology Laboratory, Canada A female Canadian citizen staying on the ninth floor of the Metropole Hotel in Hong Kong returned ill to Toronto on February 23, 2003 and infected her son. By March 14, 2003, the woman and her son had died, and three additional family members had become ill. Thus began the SARS epidemic in Canada. A total of 438 cases, 251 probable and 187 suspect were identified resulting in 44 deaths. Health Canada’s National Microbiology Laboratory has processed over 13,700 samples for SARS diagnostics. This presentation will review the original virus isolation, rapid diagnostic tool development, and the lessons learned in dealing with emerging pathogens in the face of an epidemic situation. 11:30 Case Study: Detection and Response to Microbiological Threats Speaker to be confirmed 12:10 Concluding Discussion All workshop speakers available to take questions 12:30 End of Workshop Main Conference Monday, December 8, 2003 1:00 Registration, Exhibit/Poster Set-up, Refreshments 1:55 Chairperson’s Opening Remarks Steve Semancik, Ph.D., Physicist and Project Leader of the Chemical Microsensor Program at NIST, Chemical Science and Technology Laboratory, NIST Array Technology 2:00 The Array Biosensor for Simultaneous Detection of Multiple Biological Warfare Agents Tammy A. Santana, Ph.D., Principal Biochemist, Constellation Technology Corporation Constellation Technology and the Naval Research Laboratory have developed the Array Biosensor, an immunosensor as sensitive as the ELISA. Current prototypes consist of: 1) an integrated point detector containing automated fluidics and optics and an on-board processor, and 2) an integrated standoff detector with all the features of the point detector and two types of aerosol collectors. On both prototypes, multiple assays are performed in parallel on multiple samples in about 10 minutes. 2:30 Application of Microarray Technology and Bioinformatics to Pathogen Surveillance and Diagnosis of Respiratory Illness David A. Stenger, Ph.D., CBMSE, Naval Research Lab and Epidemic Outbreak Surveillance (EOS) Program, U.S. Air Force Surgeon, General Directorate of Modernization (SGR)DNA microarray technology offers considerable promise for the detailed molecular characterization of pathogens and their effects on their hosts. A series of individual projects aimed at these applications will be described, along with a critical assessment of the technical hurdles that must be overcome. Mass Spectrometry 3:00 Bioinformatics-Based Strategies for Rapid Microorganism Identification by Mass Spectrometry Plamen A. Demirev, Ph.D., Research Scientist, Applied Physics Laboratory, Johns Hopkins University Approaches for microorganism identification, exploiting the wealth of information contained in prokaryotic genome databases, will be discussed. They are based on matching sequence-derived masses against experimentally-determined masses of biomarker proteins. Statistical analysis of proteome uniqueness provides a means to evaluate the probability of false identifications. The specificity can be improved multifold by: a) imposing rational constraints on the number of potential biomarkers; b) accounting for the most-common post-translational modifications; c) gene sequence analysis to create in silico protein biomarker databases. 3:30 Refreshment Break, Exhibit/Poster Viewing 4:00 MALDI Mass Spectrometry as a Screening Tool for Microorganism Identification Karen L. Wahl, Senior Research Scientist, Pacific Northwest National Laboratory Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS) is an analytical tool readily amenable to high throughput screening for a variety of microorganisms. Pacific Northwest National Laboratory (PNNL) has established a protocol to generate a library of signature spectra from the microorganisms based on the reproducible ions observed from replicate cultures and replicate MALDI-MS spectra collected. This includes the development of statistical algorithms to automate peak extraction, signature generation and ultimately unknown identification based on the signature library. Simple microbial mixtures, samples cultured under different laboratory conditions, and data from other laboratories have successfully been identified by the automated algorithms. An overview of our development of MALDI-MS for rapid screening and identification of microorganisms will be presented. 4:30 PCR and Protein Markers for Mass Spectrometry-Based Biodetection Alvin Fox, Ph.D., Professor, Dept. Pathology & Microbiology, University of South Carolina, School of Medicine Genes and their expressed proteins both provide logical targets for biodetection. Real-time PCR currently represents the gold-standard in terms of specificity and sensitivity. However, in environmental samples which contain complex microbial populations (many closely related to Bacillus anthracis) state-of-the-art PCR-mass spectrometry offers the additional advantage of identification by accurate molecular weight determination (mass spectometry) and sequence (tandem mass spectrometry). Protein detection, ultimately, may be simpler to implement in the field, and glycoproteins recently identified in the exosporium might serve as useful targets. 5:00 Bacteriophage Amplification in Conjunction with Analytical Chemical Methods Kent J. Voorhees, Ph.D., Professor, Department of Chemistry and Geochemistry, Colorado School of Mines Bacterial phages are known to specifically infect bacteria and rapidly multiply to form thousands of copies. This selective biological process has been used to lower the whole cell bacterial MALDI-TOF-MS detection limit from 107 to 100 cells/mL by employing the bacterial phages to produce a new amplified marker (biomarker amplification) indicative of the target bacterial cell. 5:30 End of Day One Tuesday, December 9, 2003 8:00 Exhibit/Poster Viewing, Coffee and Pastries 8:55 Chairperson’s Opening Remarks Alvin Fox, Ph.D., Professor Dept. Pathology & Microbiology, University of South Carolina, School of Medicine 9:00 BioDefense R&D in the Federal Budget for 2004 Kei Koizumi, Director, R&D Budget and Policy Program, American Association for the Advancement of Science Since the terrorist attacks of fall 2001, U.S. Government spending on counter-terrorism activities has grown dramatically. In particular, federal spending on biodefense R&D has quintupled to nearly $2 billion a year. The National Institutes of Health and the new Department of Homeland Security are responsible for a growing, diverse portfolio of research on all aspects of biodefense, including detection, fundamental science, countermeasures, and rapid response. The presentation will review the federal investment in biodefense R&D for 2004. Sensor Technology 9:30 Solid State Microsensors For Detecting Chemical Warfare Agents Steve Semancik, Ph.D., Physicist and Project Leader of the Chemical Microsensor Program at NIST, Chemical Science and Technology Laboratory, NIST We describe MEMS-based solid state microsensors that can produce distinctive responses to simulant molecules, and to agents such as sulfur mustard (HD), sarin (GB) and tabun (GA) in the ~10 ppb (10 nanomole/mole) concentration range. The ~ 100 µm x 100 µm devices rely upon adsorption-induced changes in electrical conductance at the surfaces of nanocrystalline oxide thin films for detecting hazardous compounds. We illustrate the importance of using both (rapid) temperature programming within multielement arrays and signal processing techniques for recognition and quantification of target analytes. Inherent advantages of the microsensor technology being developed include small size, low power consumption, and compatibility with on-chip electronics. 10:00 CANARY B-Cell Sensor for Rapid Identification of Pathogens Mark Hollis, Leader of the Biosensor and Molecular Technologies Group, MIT Lincoln Laboratories A novel type of biosensor for rapid pathogen identification is being developed that uses B cells, or white blood cells, as the sensing elements. B cells, which function within the body to detect pathogens, respond in a fashion that is much more rapid, sensitive, and specific than currently vailable man-made sensors. Our sensor cells are genetically engineered to selectively bind to specific pathogens and then emit photons starting in less than a second after the pathogen is bound. A biochemical signaling mechanism inside the cells provides a response that enables detection and identification of even a few pathogenic particles. The rapid 3 minutes identification of pathogens that this technology enables should be particularly important for biological-warfare agent detection, medical diagnostics, food safety assurance, environmental monitoring, and other applications. 10:30 Refreshement Break, Exhibit/Poster Viewing 11:00 Portable Integrated Biological Threat Detection System Peter Andreotti, Ph.D., President and Chief Science Officer, ASD BioSystems, Inc. ASD BioSystems has been addressing the problem of rapid biothreat agent detection by developing a man-portable integrated system for collection, processing, tracking, immunoassay, and reporting for environmental swab, wipe, filter, and liquid samples. This presentation will focus on the sample collection and processing components, hardware, software and portable luminescence immunoassay technologies which have been integrated into the hardened and decontaminable system. 11:30 Surface Plasmon-Coupled Emission: Ultra Sensitive Fluorescence Detection Technology Zygmunt Gryczynski, Ph.D., CFS Assistant Director, Associate Professor, School of Medicine Center for Fluorescence Spectroscopy, Dept. of Biochemistry and Molecular Biology, University of Maryland Fluorescence technology is the foundation of numerous analyses in sensing, medical diagnostics, biotechnology and gene expression. We present a new generic technology which will improve sensitivity 1000-fold, suppress unwanted autofluorescence from the sample and is easily implemented in many detection device formats. This technology, surface plasmon-coupled emission (SPCE) is in essence the reverse process of surface plasmon resonance (SPR), which is the absorption of light by a metal film at a specific angle of incidence. 12:00 Luncheon Sponsored by The Knowledge Foundation 1:25 Chairperson’s Remarks Peter Andreotti, Ph.D., President and Chief Science Officer, ASD BioSystems, Inc. RT PCR 1:30 From Real-Time PCR to Polymeric Biosensor Detection Without Amplification Michel G. Bergeron, MD, Head of the Infectious Disease Research Center of Laval University, Canada We have developed the first FDA approved Real-time PCR diagnostic test, now commercialized for the detection and identification of Group B Streptococci in pregnant women at delivery. Several assays using this technology are in the pipeline. However, most real-time PCR assays rely on FRET and require covalent labelling of nucleic acid probes. We recently reported a novel polythiophene sensor able to transduce nucleic acid hybridization into optical and electrical signals. This versatile detection method does not require any chemical reaction or amplification of the analyte and can detect and identify a few hundreds molecules of DNA in minutes. 2:00 The Use of Portable Real-Time RT-PCR Technology for the Rapid Identification of Selected Viruses Affecting Food Animals Johnny Callahan, MS, MT(ASCP), Scientist - Viral Assay Development, Tetracore, Inc. The U. S. Department of Agriculture and law enforcement officials consider the agricultural sector is vulnerable to economically devastating biological threat agents. Research efforts are now focused on the logistical considerations necessary to confront the problem of detecting high-consequence pathogens outside the comfort of a reference laboratory. To address these problems much work has been done towards the design and development of rapid, portable real-time PCR tests and supporting equipment as a means to provide fast, accurate answers wherever the presence of a high-consequence pathogen is suspected. Potentials threat agents, available tests and portable equipment options will be discussed. 2:30 Enhancement of PCR and Biosensor Based Bloodborne Pathogen and Toxin Detection Systems Matthew Ewart, MS CT (ASCP), Project Principle Investigator, Center for Biological Defense, University of South Florida Human blood is a living sample matrix that begins changing at the time of removal from peripheral circulation and remains a challenging material from which to efficiently extract analyte. Levels of bacteremia associated with common bacterial pathogens in adults and children may be less than 1 CFU per ml. The use of low volume sample inputs for blood culture is known to diminish detectability of bacteremia in adults. Utility for nucleic acid and biosensor based detection systems may well be enhanced by efficient bloodborne pathogen concentration technologies. We have developed a large volume blood sample processing system that provides selective solublization of human blood elements while facilitating analyte extraction in automated and fieldable devices. 3:00 Intelligence Through Sensor Data Fusion, Interoperability, and Standard Sensor Data Processing Platforms Joshua Pressnell, Senior Software Engineer, RTI Practical engineering aspects of the next generation of COTS Digital Critical Infrastructure (DCI) integration products, building blocks, components, and subsystems are reviewed with the focus on the domain of Chemical, Biological, Radiological, Nuclear, and Explosive (CBRNE) sensors and C4ISR and/or Consequence Management systems and infrastructures. A point of view is presented that the only way to achieve true interoperability across sensors and systems is to drive forward one or more reference implementations that solve a current problem or set of problems. These reference implementations will then need to evolve such that they can be packaged just as Ethernet cards or USB devices are currently in a Plug and Play (PnP) environment. Defacto and/or standard protocols will emerge which allow them to PnP into a common operational multi-level, multi-operator, shared environment. 3:30 Refreshment Break and Poster/Exhibit Viewing Nano Technology 4:00 Multiwalled Carbon Nanotube Electrodes for Gas Sensing Nikhil A. Koratkar, Ph.D., Assistant Professor, Mechanical, Aerospace and Nuclear Engineering, Rensselaer Polytechnic Institute This paper describes the design, development and proof-of-concept testing of a carbon nanotube ionization device for gas sensing. The sensor features multiw alled carbon nanotube electrode films separated from each other by several tens of microns in a parallel plate condensor arrangement. By exploiting the extremely high electric field in the vicinity of sharp nanotube tips the device is able to efficiently breakdown (ionize) target analytes and fingerprint the voltage and current discharges to identify the gas specie and its concentration. 4:30 ‘Smart Solid-State’ Nano-Materials for Real-Time Biodetection David Armstrong, SCCM, Executive Vice President, IatroQuest Corporation A unique photonic biosensing ‘smart’ material (Bio-Alloytm) takes advantage of the convergence of nanotechnology, advanced semiconductor materials and biotechnology. The unique attributes of Bio-Alloytm ‘solid state’ biosensing allows for a ‘detect-to-warn’ in comparison to many existing ‘detect-to-treat’ technologies. The potential applications for detection of harmful bio-agents will be presented. 5:00 Bioconjugated Nano Particles for Biotechnological Application Weihong Tan, Ph.D., Professor of Chemistry, University of Florida (Abstract unavailable at time of printing) 5:30 End of Conference AbstractAs the world increases its focus on identifying detrimental agents, the need for quick and accurate assessments of biological and chemical compounds has become a focal point within the worldwide detection and identification community.Get the most recent information from our international group of experts to learn and discuss the present and future aspects of detection technologies. This conference explores the latest research and state-of-the-art technologies of biological and chemical detection within this rapidly advancing field of science. Get Full Details About This Report >> |
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