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With an emphasis on rapid detection and low-cost identification, this publication provides solutions to the challenges presented by the evolving pattern of complex biological threat agents of various origins.
Completely updated with the latest information, including narratives, charts, graphs and question & answers, this essential reference tool addresses the following important topics:
- PCR and Non-PCR Based Detection
- Reagent and Reagentless Based Detection
- Point-of-Care Toxin & Pathogen Detection
- Utilizing Bacteriophage for Biological Threat Response
- Real-World Sample Preparation for Toxin & Pathogen Detection
Table of Content
Chapter 1
The Neccesity for Improving Biodetection Technologies
Robert Hooks, Deputy Assistant Secretary for WMD and Biodefense, U.S. Department of Homeland Security
In the biodefense area, DHS efforts focus on providing leadership by coordinating with other Federal partners, state/local/
Chapter 2
SAFE: Sequencing for Avian Flu Epidemic
Niveen Mulholland, PhD, Senior Scientist, MRI-NCR, Midwest Research Institute*
We have developed a system for detecting mutations that would give rise to potential pandemic-causing influenza strains. The system, SAFE: Sequencing for Avian Flu Epidemic, first uses real time RT-PCR to detect H5, the highly pathogenic avian influenza most likely to cause a pandemic. We next use pyrosequencing to detect codon changes encoding amino acids known to define human versus avian influenza signatures. The SAFE real time RT-PCR assay specifically detects H5 in a multiplex reaction designed to detect a region of the Matrix gene common to all Influenza A subtypes. The H5 primer/probe set is specific; it does not cross react with other Influenza A subtypes tested. The M primer/probe set serves as an internal control, detecting all subtypes tested. Pyrosequencing assays were developed to screen, at the nucleotide level, for 52 amino acids changes defined by Chen et al (2006) to be avian- or human- specific. A library has been built to screen the sequence data generated and properly identify the strain in question as a potential threat. This surveillance system described here will allow the global community to monitor for H5N1 and for mutations that will render the virus more infective and virulent to humans. *In collaboration with: N.Waybright, E.Petrangelo, P.Lowary & J.Bogan
Chapter 3
Next Generation Automated Multi-Target Detection Platform for Closer to Source Diagnostics
Todd Ritter, CEO of Applied Science & Technology, Idaho Technology Inc.
Idaho Technology has developed a highly multiplexed detection system capable of concurrently identifying and genetically discriminating dozens of viruses and bacteria. The syringe-loaded system utilizes a flexible plastic pouch combining automated sample preparation, reverse transcription for RNA viruses, and two-stage nested multiplex PCR performing 120 discreet analyses simultaneously. Capable of processing a variety of sample types, the small, lightweight diagnostic system represents a next generation in automated detection systems.
Chapter 4
Rapid Multiplexed Nucleic Acid and Antibody Based Sensor for Biothreat Detection
Michael R. Meyer, Director of Laboratories, ICx Biosystems
RapidPlex is a fully automated, triggered confirmation system designed for detection and identification of bacteria, virus, and toxin threats in 10 minutes. The RapidPlex system provides simultaneous, multiplexed detection of protein and DNA/RNA markers through parallel antibody and nucleic acid-based assays. Antibody-based detection utilizes multiplexed sandwich assays on spectrally encoded microspheres followed by high resolution imaging of individual beads for detection of spores/cells captured on the bead surfaces as well as detection of toxins and viruses coating the bead surfaces. The nucleic acid-based process includes automated cell lysis and purification, followed by multiplexed DNA/RNA amplification and detection of the DNA amplicons on spectrally encoded microspheres.
Chapter 5
Multiplex Detection of Biothreat Agents by Fluidic Force Discrimination
Gary W. Long, PhD, Vice President and Senior Scientist, Tetracore, Inc.
Fluidic Force discrimination (FFD) has recently been applied to the detection of Biothreat agents. In this talk, we will describe the multiplex detection of 3 or more agents by an immunoassay developed for FFD, and demonstrate that sensitivity of detection is similar or greater than that obtained in lateral flow devices and by ELISA. We will also describe improvements to the sensitivity of detection of nucleic acids using this technology.
Chapter 6
Use of CANARY™ for Rapid, Automated Collection and Analysis of Bioaerosol Samples
Thomas Hazel, PhD, Vice President Research, Innovative Biosensors, Inc.
CANARY™ is a cell-based technology that enables rapid identification of bacterial, viral, and toxin targets in liquid or aerosol samples. In this presentation we describe the testing and validation of our new BioFlash™ instrumentation, designed to provide ‘detect-to-
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