12:00 pm Registration for Pre-Conference Workshop

1:00 Welcome by Workshop Chairperson

1:55 Silicon Microstructures: A New Approach for Molecular Biology Applications
Marie Archer, Ph.D., Post Doctoral Fellow, CBMSE, U.S. Naval Research Laboratory
The rapid identification of harmful organisms has become an important priority in various fields such as clinical diagnosis and environmental surveillance. Since each organism poses a particular genetic signature they can be identified by their nucleic acid sequence (DNA and/or RNA). For this purpose the nucleic acids have to be extracted from a cell lysate using a solid phase before any further downstream process. Recently there has been an increasing demand for portable devices to perform this type of assay and therefore solid phases with integration capabilities are highly desirable. Their development requires an interdisciplinary effort between materials science, chemistry and molecular biology to address the variables that affect the performance. Silicon is an ideal candidate for this purpose due to its microfabrication properties and the flexibility to chemically functionalize its surface. This talk will discuss the development and characterization of non-selective and selective solid phases fabricated with silicon microstructures. Non-selective solid phases are used to separate the nucleic acids (DNA and RNA) from other components present in their medium that can interfere with downstream processes. This separation is not based on the nucleic acid sequence but rather in their intrinsic electric charge which allows them to interact electrostatically with a charged surface. Selective solid phases are used to separate a particular nucleic acid from a population containing various types. The separation is based on the recognition of one nucleic acid sequence, used as a probe, to its matching partner or complementary sequence. An important variable in either of these two types of solid phases is the surface area which has to be large to capture enough quantities of the nucleic acids. The geometry, the feature size and the surface roughness are particularly relevant for selective solid phases. All these variables can be controlled by fabricating solid supports through silicon micromachining. Silicon has the additional advantage of allowing various types of chemical modifications through which the surface properties can be tailored for a specific application.

2:35 Optimization and Application of FTA-Based Technology for DNA/RNA Purification
Michael Harvey, Ph.D., Director of Development, Microarrays and Molecular Biology, Whatman Inc.
The collection and preparation of clinical samples for molecular analysis is important for expanding applications in genetic identification, drug discovery, predictive medicine and pharmacogenomics. To simplify sample processing we have developed two chemically treated devices which are useful for dried clinical sample collection, sample archiving and, most importantly, DNA preparation for amplification. Nucleic acid preparation from samples collected on a treated matrix is simple, rapid and automatable. Blood and buccal cell samples collected on FTA®, a surfactant modified matrix, and FTA® Elute, a chaotropic salt modified matrix, can be stored for over 10 years under ambient conditions as demonstrated by STR analysis. We have demonstrated the use of DNA from these samples for genetic identification, real time PCR, DNA sequencing, and allele specific hybridization methods. Increasing demand for nucleic acids from archived samples dictates that systems and devices should be able to support whole genomic amplification. We have examined the recovery of DNA from samples archived on treated matrices and evaluated its’ suitability for whole genomic amplification. We have also measured the inter-sample variability and correlated this with nucleated cell counts, hematocrit and sample age. It is clear that blood and buccal samples dried on chemically modified matrices are stable and provide an excellent source of nucleic acids for future studies. The collection and storage of samples for RNA analysis is becoming an important application in pharmacogenomics. The stability and integrity of isolated RNA populations is essential for accurate analysis. Samples of tissue culture cells collected on FTA can be used to purify RNA populations which support RT PCR and real time analysis. 

3:15 Networking Refreshment Break

* Separate Registration Required

4:25 Development of Portable, Microfluidic POCT Devices for Nucleic Acid-Based Detection
Nick Parham, Ph.D. EDM, Laval University
Nucleic acid (NA)-based technology allows the development of tests with a controllable, yet broad range of specificity. For example, the presence of a diverse range of sepsis-causing microorganisms in blood can be detected by PCR-based tests. Similarly, specific single point-mutations (SNPs) causing genetic diseases can be identified. For such tests, problems such as PCR inhibitors, low target NA copy number and high background NA concentration can often confound results. These problems have largely been resolved for macro-scale samples using various NA purification techniques. However, low sensitivity remains as an important obstacle for the detection of rare targets such as water-based pathogens or sepsis-causing microbes. Another problem associated with NA-based testing is that it often requires large and expensive equipment and highly trained operators. As such, samples are sent to specialist laboratories for processing, which can involve significant time delays and costs. Portable total analysis systems can facilitate environmental monitoring and point-of-care testing. Moreover, low-cost, disposable systems that contain stable reagents will make these technologies available in low-resource settings. We are developing compact disc (CD)-based microfluidic systems for NA extraction, purification and detection from clinical samples for various applications in the molecular diagnosis of infectious diseases. Our ultimate goal is to produce low-cost, disposable microfluidic diagnostic CDs that can be operated in a portable device for point-of-care diagnostics.

5:05 Close of Pre-Conference Workshop