Detection Technologies

8:00 Registration, Exhibit Viewing/Poster Setup, Coffee and Pastries

8:45 Current Department of Homeland Security Initiatives in Bioforensics and Biodetection
David R. Hodge, PhD, Program Manager, Chem-Bioforensics, Chem & Bio Division, Science & Technology Directorate, U.S. Department of Homeland Security
Abstract will be available shortly. Please come back for the latest updates on the Program.

INTEGRATED SAMPLE PROCESSING AND MULTIPLEXED DETECTION SYSTEMS

9:15 The NRL cBASS®: Using Magnetic Microbead Labels for Attomolar Molecular Assays in Complex Matrices
Lloyd J. Whitman, PhD, Head, Code 6177-Surface Nanoscience and Sensor Technology Section, Naval Research Laboratory
The Naval Research Laboratory’s compact Bead Array Sensor System (cBASS®) is under advanced development for multiplexed analysis of complex samples. The portable prototype includes a suite of technologies for microfluidic, magnetic microbead-based microarray assays performed on a magnetoelectronic sensor chip. A key aspect of the system is a fluidic force discrimination assay, whereby analytes are captured, labeled with microbeads, and nonspecifically bound beads are removed by controlled fluidic forces. We have achieved multiplexed toxin detection at attomolar concentrations in <20 min in a variety of complex matrices. We have also demonstrated detection of bacterial DNA without PCR.

9:45 Multiplexed Biological Testing for Diagnostics and Defense
John C. Carrano, PhD, Vice President, Research and Development, Luminex Corporation
During the past several years, we have witnessed an explosive growth in the development of rather sophisticated biological detection technologies. This has been driven, in part, by at least two major needs. The first is the desire to greatly reduce healthcare costs by developing simple to use, yet accurate, point-of-care diagnostic instruments which would allow a clinician to rapidly diagnose, and prescribe treatment for a wide variety of diseases. The second driver is the unfortunate and growing threat of bio-terrorism. Hence the motivation to pursue a unified technological approach to point-of-care diagnostics and biological sensing. In this talk we will present novel technological approaches using bead-based multiplexing of biological assays to address the compelling, yet at times seemingly contradictory requirements for the development of advanced products for both biological agent sensing and point-of-care diagnostics.

10:15 Two New Multiplex Assay Panels for the Detection of Vesicular Viruses in Agricultural Samples
Pejman Naraghi-Arani, PhD, Deputy Associate Program Leader - Assays, CB Division, Nonproliferation, Homeland and International Security, Lawrence Livermore National Laboratory
Over the past three years, scientists in the Chem/Bio Division at Lawrence Livermore National Laboratory have developed four multiplex assay panels based on the Luminex liquid bead technology. These panels have included assays for the detection of both bacteria and viruses from complex sample matrices and in multi-lab tests have proven reliable and highly sensitive and selective. Here we present data from our two latest assay panels both targeting viruses that cause vesicular symptoms in cows and pigs.

10:45 Refreshment Break, Exhibit/Poster Viewing

EMERGING PATHOGEN DETECTION TECHNOLOGIES

11:15 Advances in Electronic Microarray Technology
Dalibor Hodko, PhD, Director, Advanced Technology, Nanogen, Inc.
An overview of the latest developments and applications of the electronic microarray technology will be given, in particular with respect to on-chip amplification and miniaturization of the platform. The presentation will for the first time demonstrate real-time detection of DNA amplification in an array format. Recent efforts toward integrating sample preparation with the electronic microarray platform will be summarized.

11:45 Resequencing Pathogen Microarrays (RPM) for Simultaneous Detection and Definitive Identification
Clark Tibbetts, PhD, Executive VP & CTO,
Klaus Schafer, MD, MPH, President & CEO, and
Matthew Lorence, PhD, MBA, Senior VP, TessArae Inc.
TessArae’s Resequencing Pathogen Microarray (RPM) simultaneously detects and identifies hundreds of pathogens. RPM results are gene sequences of detected pathogen(s) that distinguish known and previously unknown variants. A clinical study in 2005 demonstrated superior sensitivity and specificity, with no false positives, compared to benchmark microbial cultures and PCR tests. Within three months thirteen variant H3N2 influenza strains were identified. RPM provides single-specimen, single-test, same-day results for real-time global epidemiology.

12:15 Biodetection of Aerosolized BWA: Challenges and Pitfalls
Alex Sabelnikov, ScD, PhD, MD, Deputy Director, Center for Security Studies and Research, East Carolina University
In spite of a recent and spectacular progress in the research and development of real-time biosensors, several challenges still remain to be addressed. One concerns the sensitivity and selectivity of a biosensor. What airborne BWA concentrations should the sensors really detect in view of the agent's infection potential? How do the current sensors do in this respect? Is the further increase in sensitivity absolutely required? These and other questions are the subject of the presentation.

12:45 Luncheon Sponsored by The Knowledge Foundation Technology Commercialization Alliance

MULTI-RESOLUTION PATHOGEN TOXIN DETECTION AND TRACKING

2:00 Integrated Genomic Barcoding System for Detection and Therapeutics of First, Second and Third Generation Biothreats
Francis Larson, PhD, Maricel Kahn, PhD, and Willy Valdivia-Granda, PhD, CEO, Orion Integrated Biosciences Inc.
The developments on genetic engineering and the proliferation of synthetic biology techniques makes possible to produce de novo natural infectious agents and develop a new generation of chimeric and synthetic bioweapons that could render current detection, diagnostics, prophylactic and therapeutic countermeasures infective. To address these challenges, we will present the current progress in the application of genomic barcoding technology and its use in development of interoperable and standardized pathogen detection and genomic diversity assessment tools. Benchmarking results including sensitivity and coverage analysis will be presented and discussed. The implications of our work in identification and tracking of both known and unknown bioagents and the development of therapeutic countermeasures will be summarized.

2:30 Nanoplex™ Biotags: Robust, Multiplexed Optical Detection Labels for Ultrasensitive, No-Wash Assays
Sharron G. Penn, PhD, Chief Operating Officer, Oxonica Inc.
Oxonica has developed a novel type of nanoparticulate optical detection label based on surface enhanced Raman scattering (SERS). The physical, chemical, and optical properties of these novel nanomaterials will be described, and their performance in two assay formats described. Nanoplex™ Rapid is a multiplexed, quantitative, lateral flow immunoassay with excellent sensitivity. Nanoplex™ Direct assay is a multiplexable, homogeneous, no-wash assay. We will present exciting results from both platforms for the detection of infectious disease agents.

3:00 Magnetic Labeled Biosensor for Point of Care Diagnostics
Thea van der Wijk, PhD, Senior Scientist, Molecular Diagnostics, Philips Research, The Netherlands
Philips Research is developing a handheld biosensor for the detection of analytes in body fluids. Superparamagnetic nanoparticles are used as labels, which allow for fast, single step assay formats without any fluid wash steps and detection in raw samples (e.g. blood and saliva). A one minute competition assay has been demonstrated for the detection of single-epitope molecules, and picomolar sensitivity has been reached for assays with a sandwich-type format.

3:30 Glycoconjugate Based Synthetic Biomarkers for the Precise Detection of Toxins and Pathogens
Suri S. Iyer, PhD, Assistant Professor, Dept of Chemistry, University of Cincinnati
Antibodies are the recognition molecules of choice for various biosensing applications. However, there is a great need to develop robust, inexpensive synthetic biomarkers that are amenable to scale up and require no refrigeration. We have developed tailored glycoconjugates that exhibit antibody-like selectivity and sensitivity. These versatile synthetic molecules can be readily incorporated onto existing biosensor platforms. The synthesis and application of these recognition elements is the focus of our presentation.

4:00 Refreshment Break, Exhibit/Poster Viewing

PATHOGEN DETECTION AND CLASSIFICATION IN ENVIRONMENTAL SAMPLES

4:30 Rapid Determination of Bacterial Load for the Assessment of Water Quality
Rolf A. Deininger, PhD, Professor, and Ji Young Lee, PhD, School of Public Health, The University of Michigan
The current methods for assessing water quality include: Heterotrophic plate count (HPC), acridine orange count (AODC), direct viable count (DVC), fecal coliform count and others. These methods take from 1 - 7 days to obtain results. There remains a need for rapid, on-site evaluation of the bacterial quality of water. A rapid luminescence based system has been developed that can effectively measure generic total bacteria count (TBC) and, with the utilization of specific antibodies, identify generic E.coli. The TBC can be available within 10 minutes with a selection limit of less than 1000 cells. The study compares the luminescence field system with standard methods. The equipment is portable and the reagents are field stable. The system is suitable for laboratories, food processing plants as well as farms.

5:00 Biodetection: Identifying and Classifying Knowns and Unknowns within a Complex Mixture
Anthony P. Caruso, Vice President, febit inc.
In many applications it is necessary to identify organisms within a complex, undefined mixture, such as samples from blood, soil, agricultural sources, air samples and so on. We present a proprietary approach that utilizes our microarray technology and a combination of computational algorithms, carefully chosen biomarkers, decision support trees and graphs, which allows for the comprehensive, high-throughput identification of specific species within a mixture, and a mechanism to help identify and classify unknown organisms that have not yet been sequenced. In the area of biodefense, such an approach allows for quick identification of known pathogenic organisms, and the ability to alert when novel ones are detected.

5:30 A Powerful New Device and Method for Detecting and Concentrating Nucleic Acids from Complex and Dilute Samples
Andre Marziali, PhD, Director, Engineering Physics, Dept of Physics and Astronomy, University of British Columbia
Canada Purification of nucleic acids from complex and dilute samples remains a challenging task for many applications, including pathogen detection, environmental samples, and recovery of high molecular weight DNA. We have developed a novel electrophoretic technology for efficiently purifying and concentrating even very low levels of nucleic acids from samples where the majority of existing isolation techniques fail. We demonstrate direct extraction of DNA from soil and other environmental samples, recovery of high molecular weight DNA, and detection of DNA at zeptomolar concentrations.
*In collaboration with: J.Pel, D.Broemeling, R.Attfield, S.Carayon, I.Chan, C.Cowdell, J.Hale, E.Holtham, S.Inglis, K.Lu, G.Mercer, V.Miao, N.Shah, G.Shibahara, L.Whitehead, J.Davies

6:00 Concluding Discussion, End of Day One

Friday, November 2, 2007

8:00 Exhibit/Poster Viewing, Coffee and Pastries

POINT-OF-CARE AND REMOTE PATHOGEN AND TOXIN DETECTION SYSTEMS

8:45 Modifications to Botulinum Protease within Neurons
Jon P. DeGnore, PhD, Director of Proteomics and Mass Spectrometry, Dept of Physiology, Tufts University
Because of their high potency, ease of production and difficulty for detection, Botulinum neurotoxins (BoNT) have been classified as a Category A bioterrorist threat. In the case of botulism antidote drug development, the BoNT light chain protease is the drug target, yet this protein remains poorly understood in several critical ways. Mass spectrometric data has helped us characterize the BoNT/A light chain. This result was critical in our drug discovery efforts as we have shown that variations in the carboxyl end of LC can seriously affect the response to inhibitor.

9:15 Portable Integrated Battlespace Biodetector
Martin B. McDonnell, PhD, Principal Scientist, Defense Science and Technology Laboratory, Porton Down, United Kingdom
This presentation will describe the development of a prototype autonomous BW agent detection and identification system with the aim of achieving a similar capability to currently deployed vehicle mounted systems but in a smaller, man portable unit. The system has involved the modification and integration of relatively mature collection and sensor technologies, combined with sample processing and automated liquid handling, to produce a modular system. The aerosol collector is the Horizontal Wetted Wall Cyclone that supplies a continuous liquid sample for the Light Scattering Surface Plasmon Resonance (LS-SPR) biosensor - a low logistics burden technology which provides near real-time identification of the full range of BW agents by a combination of refractive index and light scattering detection using a single disposable multi-analyte sensor surface.

9:45 Directed Self-Assembly Fabrication of BioChemsensor Devices from Nanoparticles
Michael J. Heller, PhD, Professor, Depts of Bioengineering / Electrical & Computer Engineering, University of California San Diego
Our work is focused on assembling “micron size” biosensor devices from luminescent, fluorescent and enzyme derivatized nanoparticles. The structure for a nanolayered glucose sensor device includes a base layer of biotin/streptavidin nanoparticles, a layer of glucose oxidase derivatized nanoparticles, a layer of peroxidase derivatized nanoparticles, a layer of quantum dots, and a final layer of biotin/streptavidin nanoparticles. The mechanism of action involves using glucose in solution as the triggering mechanism for a final red fluorescent emission from the quantum dots. Such a device will serve as a prototype for a wide variety of applications which includes other biosensor devices, lab-on a-chip devices, in-vivo drug delivery systems and “micron size” dispersible bio/chem sensors for environmental, military and homeland security applications.

10:15 Portable Noninvasive Bacterial Spore Detection
Lou Reinisch, PhD, Principal Scientist, Veritide Ltd., New Zealand
Veritide is introducing a detector for bacterial spores that uses no reagents and is based upon fluorescence. Designed for the first responder, the detector can quickly analyze a white powder spill as likely to be anthrax or a hoax. The portable detector is one-button simple and requires about 3 minutes to measure a sample. Samples are collected on a swab and the swab is optically probed. The detector can quickly and accurately discriminate if the white powder contains bacterial spores, like anthrax, or a hoax substance. The patented technology combines reagentless photochemistry with the fluorescence signature to assure the highest level of specificity and sensitivity. The detector will have a lower detection limit of 100 spores. The detector does not destroy the sample, thus allowing further testing.

10:45 Refreshment Break, Exhibit/Poster Viewing

11:15 Rapid Isothermal Nucleic Acid Assays for the Detection of Pathogens
Andrew P. Miller, PhD, Vice President, Research & Development, Ionian Technologies, Inc.
Ionian’s rapid isothermal DNA and RNA amplification and detection technologies allow for unparalleled detection of pathogens in 5-10 minutes. The sensitivity and specificity of DNA/RNA amplification provide improved performance relative to immunoassays, allowing for detection of less than 50 genomes. Results can be read-out with a variety of methods, including fluorescence approaches and a colorimetric dipstick. Our colorimetric dipstick technology combined with a simple heating unit provides nucleic acid testing in a point-of-care format.

11:45 Multi-Center Validation Study of Multiplexed RT-PCR Assays for the Detection of Respiratory Pathogens
Sally M. Smith, Biophysicist, Chemical and Biological Countermeasures Division, Lawrence Livermore National Laboratory
In collaboration with the Ca. Dept. of Health and DoD GEIS laboratories, a validation study has been conducted for the real-time evaluation of operational performance for detecting respiratory pathogens in clinical samples. This follows and coincides with the development of multiplexed RT-PCR assays and automated instrumentation (FluIDx) for the detection of respiratory pathogens. The diagnostic panel includes multi-loci detection for potentially pandemic influenza, H5N1, seasonal influenza, parainfluenza, RSV and adenovirus. The results of this validation study show a high level of correlation with current gold-standard methods, even in the absence of sample preparation (i.e. no extraction or sample pre-concentration).

12:15 Solid State Flow Analyzer for Multiplexed Biological Testing
Adam R. Schilffarth, PhD, Optical Engineer, Luminex Corporation
Luminex Corporation’s xChip is a miniaturized, solid state flow cytometer based on Luminex’s proprietary xMAP technology and designed to be environmentally robust and have low reagent consumption. This miniaturization is made possible by integrating optics, fluidics, and electronics into a single device. The xChip analyzer will enable a true laboratory-in-a-briefcase for work outside of a controlled laboratory environment. The objective of xChip Phase 1 was to fully characterize 2-axis hydrodynamic flow in a planar microfluidic flow cell and interrogate xMAP microspheres. During Phase 1 of xChip, xMAP microspheres were successfully interrogated using a planar, microfluidic flow cell while realizing a 10-fold reduction in sheath fluid consumption relative to the current Luminex® 200™ flow analyzer.

12:45 Lunch on Your Own

ARRAY BIOSENSORS FOR TOXIN DETECTION

2:00 Nested Well Arrays for Toxin Detection
Z. Hugh Fan, PhD, Associate Professor, Interdisciplinary Microsystems Group, Biomedical Engineering, Mechanical & Aerospace Engineering, University of Florida
We have developed a method for detecting toxins that inhibit protein synthesis. Biological synthesis of proteins is implemented in a cell-free medium in an array of nested wells. To detect a toxin, a group of proteins is simultaneously synthesized in the array. The production yields of these proteins are inhibited differentially by the toxin. The toxin can thus be identified based on the unique response pattern of the array.

2:30 Whole Cell Biosensors: Chip Canaries for Toxicity Detection
Shimshon Belkin, PhD, Professor, Institute of Life Sciences, The Hebrew University of Jerusalem, Israel
Whole cell biosensors, based upon live genetically engineered cells “tailored” to sense toxic compounds, can report on general sample toxicity, avoiding the need to identify specific threat agents. We have pursued several pathways to immobilize such cells onto a solid platform and couple it into a signal transduction apparatus; these include the incorporation of bacterial reporter cells into disposable plastic biochips, and the development of a dedicated toxicity analyzer to analyze their response.

3:00 Detection of Biological Threat Contaminants in Critical Buildings
Ashok Kumar, PhD, Program Manager, Construction Engineering Research Laboratory, U.S. Army Engineer Research and Development Center
A nanoscale sensing model through Fluorescent Resonant Energy Transfer (FRET) interactions between fluorescent quantum dots (QDs) and organic quencher molecules can be used as a means of multiplexed detection of microbial contaminants in buildings. The detection of antigens can be measured through photoluminescence (PL) in solution. The multiplexing immuno-assay can provide the ability to detect several types of biological threat contaminants simultaneously in near real-time.

3:30 Refreshment Break, Exhibit/Poster Viewing

TECHNOLOGIES FOR PANDEMIC PREPAREDNESS

4:00 Identification of H5N1 Infection Biomarkers
Jose Malaga, PhD, and Willy Valdivia-Granda, PhD, CEO, Orion Integrated Biosciences Inc.;
James Long, PhD, and Carol Sabourin, PhD, Battelle Memorial Institute
H5N1 avian influenza virus, and its potential threat to human global health, has generated a renewed interest in the study of host-pathogen molecular interactions with potential use in diagnostics and therapeutics. Here, we present the temporal genome-wide transcriptional reconstruction of mice exposed to H5N1 using a new and scalable biomarker discovery tool. Our analysis revealed specific expression and repression patterns 6, 24 and 96 hours after challenge. These molecular interactions are associated with the up-regulation of genes within the cytokine-cytokine receptor, JAK-stat signaling and apoptosis pathways. H5N1 represses the expression of genes involved in complement coagulation cascades, MAPK, insulin and interferon production pathways. The reconstruction of the transcriptional network demonstrates that the H5N1 avian influenza induces specific set of biomarker genes that can be extrapolated to humans and allows its differentiation from other respiratory viral infections. The implications of this work in the development of detection and therapeutic countermeasures will be discussed.

4:30 Fast, Ultrasensitive Virus Detection
Aurel Ymeti, PhD, BioPhysical Engineering, MESA+ Institute for Nanotechnology, University of Twente, The Netherlands
We have developed an ultrasensitive sensor that could potentially be used in a handheld device to, within minutes, detect various viruses and measure their concentration. The sensor could be used to quickly screen people at airports, hospitals and emergency clinics to control outbreaks of diseases such as SARS and the bird flu. All it would take is a tiny sample of saliva, blood, or other body fluid.

5:00 A Sensitive Nanocrystal Biosensor for Rapid Detection of Pathogens
Yongcheng Liu, PhD, Senior Scientist, NN-Labs
A biosensor for detection of pathogens was developed by using CdSe/ZnS core/shell dendron-nanocrystals with high efficiency and stability as fluorescence labels and a flowing chamber with a micro-porous immunofilter. The antibody-immobilized immunofilter captured the targeted pathogens, /E. coli/ O157:H7 as an example for bacteria and Hepatitis B being a model system for viruses. The CdSe/ZnS core/shell dendron-nanocrystals were conjugated with the corresponding antibodies and then passed through the micro-porous membrane where they attached to the membrane-antigen-antibody. The efficient and stable photoluminescence (PL) of the CdSe/ZnS nanocrystals on the formed "sandwich" structure complexes (membrane-antigen-antibody conjugated with the nanocrystals) was used as the detection means. The effects of the pore size of the membranes, buffer pH, and assay time on the detection of /E. coli/ O157:H7 were investigated and optimized. The detection limit of this new biosensor was as low as 2.3 CFU/ml for /E. coli/ O157:H7 and 5ng/ml for Hepatitis B surface Ag (/HBsAg//)/. The assay time was shortened to 30 min without any enrichment and incubation.

5:30 Closing Remarks, End of Conference

· The NRL cBASS®: Using Magnetic Microbead Labels for Attomolar Molecular Assays in Complex Matrices
Lloyd J. Whitman, PhD, Naval Research Laboratory

· Multiplexed Biological Testing for Diagnostics and Defense
John C. Carrano, PhD, Luminex Corporation

· Two New Multiplex Assay Panels for the Detection of Vesicular Viruses in Agricultural Samples
Pejman Naraghi-Arani, PhD, Lawrence Livermore National Laboratory

· Advances in Electronic Microarray Technology
Dalibor Hodko, PhD, Nanogen, Inc.

· Resequencing Pathogen Microarrays (RPM) for Simultaneous Detection and Definitive Identification
Clark Tibbetts, PhD, TessArae Inc.

· Biodetection of Aerosolized BWA: Challenges and Pitfalls
Alex Sabelnikov, ScD, PhD, MD, East Carolina University

· Integrated Genomic Barcoding System for Detection and Therapeutics of First, Second and Third Generation Biothreats
Willy Valdivia-Granda, PhD, Orion Integrated Biosciences Inc.

· Nanoplex Biotags: Robust, Multiplexed Optical Detection Labels for Ultrasensitive, No-Wash Assays
Sharron G. Penn, PhD, Oxonica Inc.

· Magnetic Labeled Biosensor for Point of Care Diagnostics
Thea van der Wijk, PhD, Philips Research

· Glycoconjugate Based Synthetic Biomarkers for the Precise Detection of Toxins and Pathogens
Suri S. Iyer, PhD, University of Cincinnati

· Rapid Determination of Bacterial Load for the Assessment of Water Quality
Rolf A. Deininger, PhD, The University of Michigan

· Biodetection: Identifying and Classifying Knowns and Unknowns within a Complex Mixture
Anthony P. Caruso, febit inc.

· A Powerful New Device and Method for Detecting and Concentrating Nucleic Acids from Complex and Dilute Samples
Andre Marziali, PhD, University of British Columbia

· Modifications to Botulinum Protease within Neurons
Jon P. DeGnore, PhD, Tufts University

· Portable Integrated Battlespace Biodetector
Martin B. McDonnell, PhD, Defense Science and Technology Laboratory

· Directed Self-Assembly Fabrication of BioChemsensor Devices from Nanoparticles
Michael J. Heller, PhD, University of California San Diego

· Portable Noninvasive Bacterial Spore Detection
Lou Reinisch, PhD, Veritide Ltd.

· Rapid Isothermal Nucleic Acid Assays for the Detection of Pathogens
Andrew P. Miller, PhD, Ionian Technologies, Inc.

· Multi-Center Validation Study of Multiplexed RT- PCR Assays for the Detection of Respiratory Pathogens
Sally M. Smith, Lawrence Livermore National Laboratory

· Solid State Flow Analyzer for Multiplexed Biological Testing
Adam R. Schilffarth, PhD, Luminex Corporation

· Nested Well Arrays for Toxin Detection
Z. Hugh Fan, PhD, University of Florida

· Whole Cell Biosensors: Chip Canaries for Toxicity Detection
Shimshon Belkin, PhD, The Hebrew University of Jerusalem

· Detection of Biological Threat Contaminants in Critical Buildings
Ashok Kumar, PhD, U.S. Army Engineer Research and Development Center

· Identification of H5N1 Infection Biomarkers
Jose Malaga, PhD, Orion Integrated Biosciences Inc.

· Fast, Ultrasensitive Virus Detection
Aurel Ymeti, PhD, University of Twente

· A Sensitive Nanocrystal Biosensor for Rapid Detection of Pathogens
Yongcheng Liu, PhD, NN-Labs