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Date: 25th June, 2008

Location: Atlanta GA, USA

Summary:
Clinical - Seminar

Agenda:
Sample Preparation for Rapid Microorganisms Detection – Current Perspective and Future Challenges
George Hong, PhD, R&D Manager, Technology and Pre-Development, Millipore Corporation
Since the discovery of nucleic acids, proteins, or cellular components as targets for microbial identification, this field has advanced from culture growth-dependent verification to rapid culture-independent analysis. However, no matter how sophisticated the field has progressed, these innovative technologies often rely heavily on robust sample preparation methods to achieve consistent and accurate performance. Here, we present an overview of major sample preparation methods, which encompass collection, concentration, and purifications of the target of interest for microbial identification. Also in this presentation, we will touch on major future challenges for the advancement of rapid sample-preparation for the field of microbial identification.

Bio-aerosol Collection for Indoor and Outdoor Applications
Charles Call, PhD, CEO Biodefense, ICx Technologies, Inc.
Technologies for the collection of aerosolized pathogens and other bio-threat agents will be reviewed. Approaches which will be covered include standard and cascade impactors, rotating impactors, virtual impactors, wet-walled cyclones, and dry filters. Performance metrics for bio-aerosol collectors will be addressed. Applications of these technologies will be described in detail for both indoor and outdoor environments. Methods of characterizing the performance of aerosol collectors will be presented. The session will wrap up with a short review of approaches and products for surface sampling for pathogens.

A Water Condensation Method for the Concentration and Capture of Airborne Particles
Susanne V. Hering, PhD, Founder & President, Aerosol Dynamics Inc.
Key to the effective surveillance of airborne pathogens is the collection and delivery of these pathogens to the detection system. Generally, this requires the concentration of airborne particles from large sample airflows into a small volume of water solution. Aerodynamic concentration methods such as virtual impactors can be effective for particles from 2 µm to 10 µm, yet many bacteria are below 2 µm, and nearly all viruses are below 0.5 µm. Described here is a new method that uses condensational growth to encapsulate submicrometer and supermicrometer particles within a supermicrometer water droplet, with subsequent collection in small liquid volumes.

BEADS: A Modular Sample Preparation System for Pathogen Detection
Cindy Bruckner-Lea, PhD, Chemical & Biological Science Group Manager, National Security Directorate, Pacific Northwest National Laboratory*
Sample preparation is a limiting technology for developing fully integrated, autonomous, and portable biodetection systems. BEADS (Biodetection Enabling Analyte Delivery System) is a modular fluidics approach that bridges the gap between large volume liquid samples generated from environmental matrices (aerosol collectors, soils, water, etc.) and microscale detectors that require small, clean samples. The central components of BEADS sample preparation systems are derivatized particles, which are used for concentrating and purifying pathogens and pathogen biomarkers. The particles are automatically introduced, trapped and released in specialized flow cells. Fouling of the critical bio-interactive surface does not occur because new particles, and therefore new interactive surfaces are automatically introduced for each sample. This presentation will include a summary of sample preparation modules and results for trace detection of pathogens and toxins in complex samples and large samples (e.g., 1 liter water), an ultrasonic flow-through lysis module to increase the availability of DNA from Bacillus spores for detection, and integration of sample preparation and multiplexed detection modules.
*In collaboration with: R.Ozanich, T.Straub, and C.Warner

Sample Collection and Processing: Experiences from the Lab to the Field
Andrew Cannons, PhD, Scientific Director, University of South Florida Center for Biological Defense
Preparation and processing of samples for the detection and characterization of bioterrorism (BT) agents are essential elements for the early recognition of a BT event. This phase includes sample collection in the field to rapid processing of varied matrices in the lab followed by detection and analysis. Here we report on some of our efforts in sample preparation and processing, including sample collection training for emergency responders and processing different real life samples (food, soil, powders) for rapid pathogen characterization.

Development of an Automated Sample Preparation Device for Use on the BioSeeq®-Vet Portable PCR System
John W. Czajka, PhD, MBA, Vice President of Technology Acquisition, Smiths Detection
Smiths Detection has developed the BioSeeq®-Vet PCR System, a portable PCR instrument that performs automated sample preparation and PCR analysis. Nucleic acids are extracted and purified from samples by the Sample Preparation Unit (SPU), a single-use, self-contained sample preparation and PCR consumable. All enzymes and reagents needed by the unit are preloaded into the SPU and are temperature stable. The SPU was designed to purify DNA or RNA from a wide range of sample types and is driven by the BioSeeq®-Vet instrument.

Electric Field Driven Sample Collection, Concentration and Preparation Technologies
Kapil Pant, PhD, Manager, Microsystems and Nanosystems, CFD Research Corporation
CFD Research Corporation (CFDRC) has developed a suite of sample collection, concentration and preparation technologies that harness electric fields to implement various steps involved in integrated biodetection. CFDRC-developed technologies include (1) high efficiency, “gentle” aerosol sampling and concentration using electrostatics, (2) intelligent discrimination of target cells and spores from contaminant particles using dielectrophoresis, (3) rapid, reagent-less lysis of cells and spores using AC electric fields, (4) fast target extraction in a microsphere-based format with active, electrokinetic mixing, and (5) nano-electrokinetic sample pre-concentration for enhanced assay sensitivity.

High Affinity Chemicals Used in Sample Preparation
X. James Li, PhD, Chief Scientific Officer, Cellex, Inc.
Isolation and enrichment of detection materials (nucleic acids and proteins) from pathogens are the first and crucial step for successful detection of the pathogens. We will describe two sample preparation approaches. The first approach is a virus capture based approach; the captured virus can be directly used for downstream detection. The second approach involves isolation and enrichment of nucleic acids from highly diluted samples using a class of high affinity chemicals.

A Filtration System for Separation, Storage and Efficient Rapid Release of Plasma from Clinical Samples of Blood
Galina Fomovska, PhD, Senior Principal Scientist, Pall Lifesciences, Molecular Media R&D, Pall Corporation
We will demonstrate the effectiveness of a novel membrane system composed of asymmetric polysulfone BTS-SP 300 membrane and Accuwik® Ultra hydroxylated polyester media, for the rapid separation, collection, storage and efficient release of plasma from whole blood directly from a donor without the use of equipment. We will demonstrate that concentrations of important clinical biomolecules, lipids, proteins, and hormones, in plasma samples separated, from whole blood, stored, and reconstructed using the system are practically identical to those generated by centrifugation.

ROUND TABLE DISCUSSION:
Current Needs and Future Directions in Sample Preparation for Rapid Microbial Detection
Moderator - George Hong, PhD, Millipore Corporation