Biodetection Technologies

8:15 Registration, Exhibit/Poster Setup, Coffee and Pastries

9:00 Chairperson's Opening Remarks & Introduction
Raymond P. Mariella Jr., PhD, Director of the Center for Micro- and Nanotechnology, Lawrence Livermore National Laboratory

9:10 Opening Address
Real-Time Detection of Zoonotic Threats -
It's Time to Stop Using Taxpayers as Sentinels
Tracey McNamara, DVM, Diplomate ACVP, Midwest Research Institute
Most of the known threat agents and all of the emerging infectious diseases that have recently posed a threat to human health (Nipah, WNV, SARS, monkeypox and avian influenza) have been zoonoses. And yet, little attention has been paid to real-time detection of these infections in the animal reservoirs themselves. Although funding continues to pour into syndromic surveillance projects, baseline data on the presence of zoonoses is lacking, making it difficult to detect blips above normal background. Low cost screening assays that have been validated against a number of domestic and wild animal species must be developed if baseline is to be established.

9:40 Remote Testing for Foreign Animal Diseases Using Dried PCR Reagents and a Portable Laboratory
Johnny Callahan, MS, MT(ASCP), Scientist - Viral Assay Development, Tetracore, Inc.
Presently, testing for foreign animal diseases is performed at reference laboratories by the combined use of serologic and virus isolation techniques, supplemented by the reverse transcriptase polymerase chain reaction (RT-PCR) assay. These techniques require the availability of a dedicated laboratory facility and management of the logistic considerations associated with sample collection and transport. Portable, rapid, and easy-to-perform tests that would permit on-site diagnosis in suspected disease outbreaks would have clear advantages over present methods. The development, field experience and advantages of two dried RT-PCR PCR tests for detection of foreign animal disease viruses (FMDV and CSFV) will be presented.

10:10 Stage-Specific Diagnosis of Exposure to Biological Threat Agents
Marti Jett, PhD, Chief of Molecular Pathology Dept, Walter Reed Army Institute of Research
Use of gene response profiles to reveal phases of progression of illness has the potential to (1) provide stage-specific diagnosis and (2) enable identification of potential molecular targets for stage-appropriate therapeutic interventions for intractable illness induced by unconventional pathogenic agents. Classical methods for treatment of lethal shock as a result of pathogen exposure have targeted many inflammatory mediators of illness, yet clinical success in designing treatment regimens has been elusive. Our aim is to utilize global gene studies to better characterize the effective time frame for treatment approaches.

10:40 Refreshment Break, Exhibit/Poster Viewing

11:00 Featured Presentation
Review of State-of-the-Art Contamination Detection Technologies for Early Warning Systems for Drinking Water
Jafrul Hasan, PhD, MPH, Office of Science and Technology, Office of Water, and National Homeland Security Research Center, Office of Research and Development, U.S. Environmental Protection Agency*
Terrorist attacks have heightened concern over the potential for intentional contamination of the U.S. water system. An integrated Early Warning System (EWS) can be an important tool to ensure deployment of monitoring technologies/strategies and enhanced surveillance activities to collect, integrate, analyze, and communicate information to provide a timely warning of potential water contamination incidents and initiate response actions to minimize public health impacts. This paper will summarize the status and initial findings of a current EPA project to review the state-of-the-art technologies for integrated EWSs for drinking water, particularly for finished water supplies and distribution systems. This project outlines desired characteristics of an integrated EWS and summarizes and evaluates current and emerging EWS technologies for identifying general categories of chemical, microbial, and radiological contaminants. Specifically, it examines the state of sensor technology (including multi-parameter water quality monitors) to detect the contaminants, systems to transmit, acquire, and analyze data, sensor placement, contaminant flow modeling, alert management, data security, and protocols for notification, decision making and emergency response. The project presents conclusions and recommendations regarding future directions of EWSs including technical issues and research gaps.
*In collaboration with: J.Herrmann US EPA/NHSRC, D.Goldbloom-Helzner, ICF Consulting

11:15 Direct Detection of Select Agents in Foods Using BioVeris Technology™
Charles Young, PhD, Director of Biodefense Product Development, BioVeris Corporation
BioVeris (BV) Technology uses labels designed to emit light when electrochemically stimulated. Due to the highly selective nature of the BV detection process, BioVeris has developed a variety of tests for the detection of select agents in foods that requires little or no sample preparation. Data for the direct detection of ricin, botulinum neurotoxins, Staphylococcus aureus enterotoxins and anthrax in a variety of food matrices will be presented.

11:45 Portable High-Throughput Integrated Laboratory Identification System (PHILIS)
E. Bradley Perkins, Director of CBRN Detection Systems, EAI Corporation
The Portable High-throughput Integrated Laboratory Identification System consists of three major subsystems:
(1) Sample preparation and screening procedures;
(2) Analytical procedures laboratory;
(3) Sample storage and evidence preservation area.
We use an automated Laboratory Information Management System to manage the large number of samples. The sample processing area and the Storage operations support the PHILIS by providing scalable capability to prepare samples for the analysis process.

12:15 Dynamic Surface Generation and Imaging for the Detection of Biological Agents
Stuart A. Kushon, PhD, Manager BioDefense Group, QTL Biodefense
Dynamic Surface Generation and Imaging is presented for the detection of bioagents (Bacterial spores, Protein Toxins, etc). The mode of detection is rapid with only three minutes of incubation time required prior to the result. (A user could have results within 5 minutes even after sample preparation). Multiplexed detection of surface swabbed biological agents is presented with limits of detection at or below lethal doses, specifically anthrax, ricin, and SEB.

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

Reagentless Detection Methods

2:00 Chairperson's Introductory Review
Low Operational Cost Biodetection Systems -
A Historical Perspective and Rationale
Wolf P. Altman, PhD, Director, Program Management, Battelle
A historical perspective of the US military and homeland security efforts towards developing biodetection systems and culminating in the existing fielded systems will be presented.These systems include BIDS, JBPDS and BioWatch. While these systems provide a level of warning of biological warfare attacks, one of the main aspects of these systems that hinders their widespread usage is their operational cost. We will discuss the operational characteristics (at a very high level) of these systems and their rough order of magnitude operational costs and need to develop systems that provide comparable performance to existing systems but at significantly lower operational costs.

2:30 The Potential of LIBS for Rapid Detection and Identification of Bio-Hazards
Andrzej W. Miziolek, PhD, US Army Research Laboratory
Laser Induced Breakdown Spectroscopy (LIBS) is an emerging sensor technology with high potential for relatively low-cost and real-time detection and identification of biological threats. LIBS is based on focusing a pulsed laser on the target material and capturing the light from the laser-produced microplasma. This light is analyzed using new generation broadband high resolution spectrometers and species identification is accomplished using advanced chemometrics. Possible LIBS applications include direct monitoring of air flows (e.g. HVAC systems) as well as analysis of bio-aerosols captured on filters.

3:00 Reagentless Molecular Chemical Imaging Detection of Biothreat Agents
Patrick J. Treado, PhD, Founder and CTO, ChemImage Corporation*
Molecular chemical imaging (MCI) based on Raman and fluorescence optical spectroscopy is being applied for rapid, reagentless detection of biological threat agents in complex biological environments. The capabilities of MCI have been demonstrated in a number of government-managed trials. In this presentation, the current state of MCI technology will be assessed, as well as the results of validation studies.
*In collaboration with: M.P.Nelson, G.S.Vanni, and J.Neiss

3:30 Refreshment Break, Exhibit/Poster Viewing

4:00 Real-Time Reagentless Agent and Analyte Detection and Identification
H. James Harmon, PhD, Professor of Physics, Center for Sensors and Sensor Technology, Oklahoma State University
Reagentless detection and identification of bio- and chemical WA, pesticides, and TICs can be made optically in less than 5 seconds using a porphyrin-based monolayer. Analyte- and porphyrin-specific absorbance changes in the visible light region are measured using a hand-held spectrometer; specificity for detection of cholinesterase inhibitors including GB are made using enzyme-porphyrin complexes. LODs down to 7 ppt (0.007 ppb) can be measured. The spectrometer-detector surface package weighs less than 1 pound with a less than 6 cubic inch volume.

4:30 Real-Time Detection of Biological Aerosols with Biological Aerosol Mass Spectrometry (BAMS)
Eric E. Gard, PhD, Division Leader for Defense Biology and Bio-Detection Instrumentation, Lawrence Livermore National Laboratory
We will be discussing the use of Biological Aerosol Mass Spectrometry (BAMS) for detection of biological aerosols in real-time with no reagent consumption. The system is able to process 100,000 particles per second and can rapidly determine when a biological aerosol is present. We will also show how the system can distinguish between various biological agent aerosols in real-time.

5:00 Rapid Identification of Biological Threat Agents in Cluttered and Uncluttered Conditions Using Atmospheric Pressure MALDI Mass Spectrometry
Robert M. Serino, PhD, Director of Operations, Science & Engineering Services, Inc.*
We will describe the use of atmospheric pressure (AP) matrix-assisted laser desorption/ionization (MALDI) tandem mass spectrometry combined with proteomics and internet bio-informatics to identify real threat bioagents and threat simulants in both neat and highly cluttered conditions. The classes of agents include spores, vegetative cells, viruses and toxins, including Bacillus anthracis, Yershinia pestis, Vaccinia and Clostridium botulinum neurotoxin A studied at our company facility and at Dugway Proving Ground. The total analysis time including sample preparation is within 5-10 minutes. The data would suggest that AP/MALDI technologies offer low-cost, rapid, and effective identification of wide-ranging biological materials to afford a new and important capability.
*In collaboration with: V.M.Doroshenko, V.Berkout, N.Taranenko, G.Black, V.Laiko, H.S.Lee, and C.R.Prasad

5:30 Panel Discussion

Reagent Free Identification: Myth or Reality?

Facilitator:

Wolf P. Altman, Battelle

Panelists:

Eric E. Gard, Lawrence Livermore National Laboratory
H. James Harmon, Oklahoma State University
Andrzej W. Miziolek, US Army Research Laboratory
Robert M. Serino, Science & Engineering Services, Inc.
Patrick J. Treado, ChemImage Corporation

6:00 End of Day One

Friday, June 10, 2005

8:15 Exhibit/Poster Viewing, Coffee and Pastries

8:55 Chairperson's Remarks
H. James Harmon, PhD, Professor of Physics, Center for Sensors and Sensor Technology, Oklahoma State University

9:00 Key Note Address
Optically-Based Bio-Sensors
LTC John C. Carrano, PhD, Program Manager, DARPA/MTO
After the anthrax attacks during the fall of 2001, much focus has been placed on developing effective means to detect biological pathogens. Unfortunately, practical solutions still allude the technical community. However, DARPA's Semiconductor Ultraviolet Optical Sources program has met with success in developing deep ultraviolet laser diodes and LED's, and integrating new sources into a new generation of compact and sensitive bio-detectors. In this talk, we will describe two spectroscopic techniques useful for bio-sensing: laser-induced fluorescence, and UV resonance-enhanced Raman. We will also detail advances in optoelectronic devices that now enable these techniques to become practical as effective early-warning bio-sensors capable of continuous monitoring without consumables. In particular, we will describe innovations that have resulted in UV LED's operating at wavelengths as short as 275 nm with milliwatt class CW optical power, and laser diodes operating in the range of 340 nm to 365 nm. These devices are fabricated from epitaxial layers of AlInGaN grown on a variety of substrates. These devices are the critical components in a new class of compact, rapid, reliable, and low-cost biosensors under development in the program.

9:40 Key Note Address
Advanced Technologies for Biodetection:
From Nanosensors to Biochips
Tuan Vo-Dinh, PhD, Director, Center for Advanced Biomedical Photonics, Oak Ridge National Laboratory
This presentation provides an overview of the principle and applications of several advanced technologies developed for sensitive and selective detection of chemical and biological agents:
(1) Multifunctional biochip;
(2) Nanosensor for cell analysis;
(3) Plasmonics sensors; and
(4) Portable Raman monitor.

10:20 Refreshment Break, Exhibit/Poster Viewing

10:50 Two-Minute Detection and Identification of a BW Agent
Raymond P. Mariella Jr., PhD, Director of the Center for Micro- and Nanotechnology, University of California/Lawrence Livermore National Laboratory
The National Research Council Of The National Academies has recently released its long-awaited report "Sensor Systems for Biological Agent Attacks: Protecting Buildings and Military Bases", by the Committee on Materials and Manufacturing Processes for Advanced Sensors Board on Manufacturing and Engineering Design Division on Engineering and Physical Sciences. In this report, there are notional concepts that describe technically approaches to detect and identify a biological warfare agent or other pathogen, starting with sample collection and ending with communicating the result, in under two minutes. This talk will review important aspects of this report.
This work was performed under the auspices of the U.S. Department of Energy by University of California Lawrence Livermore National Laboratory under contract No. W-7405-Eng-48.

11:30 Universal Pathogen Identification Microarray
Joany Jackman, PhD, Senior Scientist, The Johns Hopkins Applied Physics Laboratory
Microarrays have had a great impact on the speed and accuracy of microbial identification. Universal pathogen identification microarrays have been developed that utilize ribosomal markers which can be correlated with classical microbiological strategies used for species specific identification. A pure sample derived from a bacterial colony or a semi-pure sample derived from environmental material can be analyzed in less than two hours. Viruses can be analyzed for family, type and species rapidly by this method using amplicon derived fragments. This technology is portable, lightweight (under 6 lbs) and uses few consumables.

12:00 PathAlert™, LUX™, RLS, and ChargeSwitch™: Technological Advances in Detection and DNA Extraction
Bill Folkerts, Associate Director, Biodefense, Biological Defense Systems, an Invitrogen Company
PathAlert™ and LUX™ represent novel PCR amplification approaches to detection of biological threat agents. These systems continue to advance in breadth of line and performance. Specifically, a multi-agent PathAlertª has been added to the list of COTS assays currently available. PathAlert™ assays are performed on the Agilent Bioanalyzer or ALP platforms. LUX™ assays are performed on any rtPCR commercially available platform. RLS (Resonance Light Scattering) represents a novel approach towards sensitive immuno-assays for detection of biological agents. The RLS technology lends itself to miniaturization and hand held capabilities. These technologies associated with each of these systems will be described along with performance data. ChargeSwitch™ represents a novel approach toward nucleic acid purification that does not require centrifugation or the use of ethanol in sample preparation. The technology and performance data will be presented.

12:30 Lunch on Your Own

1:55 Chairperson's Remarks
Johnny Callahan, MS, MT(ASCP), Scientist - Viral Assay Development, Tetracore, Inc.

2:00 The CT-ABS™ Arrayed Immunosensor Technology for the Simultaneous Detection of Multiple Biowarfare Agents
Tammy Spain, PhD, Principal Biochemist, Constellation Technology Corporation
The Constellation Technology Array Biosensor (CT-ABS™) is a portable, fully-automated immunosensor as sensitive as the ELISA. Multiple assays on up to 6 samples can be done in 10 minutes. Assay kits are available for Staphylococcal Enterotoxin B and Bacillus anthracis. Assay kits for Ricin and Botulinum Toxin are scheduled to be released in 2005. This presentation will discuss the performance of the CT-ABS™ as a standalone instrument and as an adaptable technology for biowarfare defense systems.

2:30 Field Detection Process Controls:
Mitigating User-Error Utilizing Off-The-Shelf PDA Interfaces
James P. Whelan, PhD, President and General Manager, Alexeter Technologies, LLC
False positive tests in biological field detection scenarios occur predominately from user-operation errors rather than actual test cross-reactivities. The development of a new optical scanning platform onto the HP4700 wireless PDA provides not only a more rapid and sensitive option to our current Guardian Reader System, but increased process controls that should minimize the occurrence of user errors. Comparison performance data and advanced functionalities will be discussed in the context of our observed user-error patterns.

3:00 Refreshment Break, Exhibit/Poster Viewing

3:15 Applications of Real-Time PCR in Pathogen Detection
Maura Barbisin, PhD, Senior Scientist, Applied Biosystems*
Fluorescent quantitative real-time PCR is used in a wide variety of molecular applications, gene expression analysis and genotyping analysis. It is also the most sensitive and accurate method for detection and measurement of pathogen levels. In this abstract we outline Applied Biosystems' strategy for the generation of real-time PCR assays that can be used to monitor different pathogens that are classified as biothreat agents. We developed a bioinformatics assay design pipeline that generates assays with
(1) near 100% PCR amplification efficiency, and
(2) a high degree of specificity for the selective amplification of a targeted gene in the presence of closely related targets.
We also describe several applications of these assays in monitoring biothreat agent levels. In particular, we show improved real-time PCR detection in the presence of various inhibitors.
*In collaboration with: O.Petrauskene, M.Tanner, M.Malicdem, M.Johnson, P.Brzoska, E.Spier, and M.Furtado

3:45 Rapid Culture - Independent Identification of Microorganisms Using One Convenient Automated Platform
George Hong, PhD, Director of Microbiology, Transgenomic, Inc.
Although many current molecular approaches allow direct detection of known microorganisms from a given sample, very few approaches are capable of detecting known and unknown microorganisms simultaneously. The automated high-throughput WAVE(R) Microbial Analysis System is one of the available technologies capable of scanning the presence of what microorganisms are present in the samples, ranging from environmental to clinical.

4:15 Genomic and Computational Analysis of Encephalic and Hemorrhagic Viruses
Willy Valdivia-Granda, PhD, Founder and CEO, Orion Integrated Biosciences Inc.
We present a comprehensive computational and experimental analysis of all sequence information available for species and strains of Flavivirus, Filoviruess, Alphaviruses and Bunyaviruses. Our results address one of the most fundamental questions about the origins of viral evolution, adaptation and divergence. The information content, preferential binding of these elements to the human MHC provides evidence of a new level of functional organization of the viral genome and have implications in the rational development of diagnostics devices, vaccines and drugs.

4:45 Selected Oral Poster Presentations, Discussion

5:15 Concluding Remarks, End of Conference