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Overview

With the emergence of new potential threats, the demand for faster, more accurate biological and chemical detection solutions continues to be a worldwide focus. Clearly defined lines of communication between developers and end users of such detectors is essential for enabling successful deployment of these technologies.

This fourth international conference in the detection series is a platform for the unprecedented exchange of information between leading academic, government and commercial researchers as they probe the latest findings within the sensor community. Gain insight into the methodologies, technical innovations and viable applications of detection technologies as our expert panel of speakers discuss such topics as:

Featuring:

• SARS Genome Sequences
• Detection and Analysis of SARS
• SARS: Ready To Return?
• Eliminating False Positive Using RT PCR
• Detecting Threat Agents - the Anthrax Example

SOLD OUT EVERY YEAR — Register today to ensure your place!

Agenda

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 Case Study: SARS - Challenges in Detection and Analysis
Speaker to be confirmed

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

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

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.

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.

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.

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 delectability 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 Refreshment Break and Poster/Exhibit Viewing

3:30 Carbon Nanotube Nanoelectrode Array as an Electronic Chip for Ultrasensitive Label-Free DNA Detection
Jun Li, Ph.D., Physical Scientist, NASA Ames Research Center
An electronic chip based on carbon nanotubes (CNTs) nanoelectrode array for rapid DNA/RNA analysis will be presented. Oligonucleotide probes can be selectively functionalized at CNT end. Ultrahigh sensitivity for detecting label-free PCR amplicon has been demonstrated using a mediator amplified guanine oxidation mechanism. The detection limit is estimated to be less than 1000 DNA molecules. This system provides a general platform for rapid molecular diagnostics in applications requiring ultrahigh sensitivity, high-degree of miniaturization, simple sample preparation, and low-cost operation.

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

Call for Posters

Industry and academic scientists are encouraged to submit poster titles for this event. One-page abstracts (8 1/2" x 11" with 1-inch margins) must be submitted no later than November 10, 2003 for inclusion in conference documentation. Additional poster submissions will be accepted until December 3, 2003 but may not be included in conference documentation.

DIMENSIONS of the poster boards are: 4 feet wide by 3 feet high
(although it could be placed vertically as well and then the dimentsions obviously would be 3'w x 4'h)

Note: If you're submitting a poster, you MUST be registered and paid registration fee plus posterboard reservation fee in advance to ensure that a posterboard is reserved for you.

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