Publications

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Recovery of Bacillus atrophaeous spores from grime-treated and clean surfaces was measured in a controlled chamber study to assess sampling method performance. Outdoor surfaces investigated by wipe and vacuum sampling methods included stainless steel, glass, marble and concrete. Bacillus atrophaeous spores were used as a surrogate for Bacillus anthracis spores in this study designed to assess whether grime-coated surfaces significantly affected surface sampling method performance when compared to clean surfaces. A series of chamber tests were carried out in which known amounts of spores were allowed to gravitationally settle onto both clean and dirty surfaces. Reference coupons were co-located with test coupons in all chamber experiments to provide a quantitative measure of initial surface concentrations of spores on all surfaces, thereby allowing sampling recovery calculations. Results from these tests, carried out under both low and high humidity conditions, show that spore recovery from grime-coated surfaces is the same as or better than spore recovery from clean surfaces. Statistically significant differences between method performance for grime-coated and clean surfaces were observed in only about half of the chamber tests conducted.

Recovery of spores from environmental surfaces is known to vary due to sampling methodology, techniques, spore size and characteristics, surface materials, and environmental conditions. A series of tests were performed to evaluate a new, validated sponge-wipe method. Specific factors evaluated were the effects of contaminant concentrations and surface materials on recovery efficiency (RE), false negative rate (FNR), limit of detection (LOD) - and the uncertainties of these quantities. Ceramic tile and stainless steel had the highest mean RE values (48.9 and 48.1%, respectively). Faux leather, vinyl tile, and painted wood had mean RE values of 30.3, 25.6, and 25.5, respectively, while plastic had the lowest mean RE (9.8%). Results show a roughly linear dependence of surface roughness on RE, where the smoothest surfaces have the highest mean RE values. REs were not influenced by the low spore concentrations tested (3 x 10{sup -3} to 1.86 CFU/cm{sup 2}). The FNR data were consistent with RE data, showing a trend of smoother surfaces resulting in higher REs and lower FNRs. Stainless steel generally had the lowest mean FNR (0.123) and plastic had the highest mean FNR (0.479). The LOD{sub 90} varied with surface material, from 0.015 CFU/cm{sup 2} on stainless steel up to 0.039 on plastic. Selecting sampling locations on the basis of surface roughness and using roughness to interpret spore recovery data can improve sampling. Further, FNR values, calculated as a function of concentration and surface material, can be used pre-sampling to calculate the numbers of samples for statistical sampling plans with desired performance, and post-sampling to calculate the confidence in characterization and clearance decisions.

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Towards developing rapid and portable diagnostics for detecting zoonotic diseases, we have developed microchip-based electrophoretic immunoassays for sensitive and rapid detection of viruses. Two types of microchip-based electrophoretic immunoassays were developed. The initial assay used open channel electrophoresis and laser-induced fluorescence detection with a labeled antibody to detect influenza virus. However, this assay did not have adequate sensitivity to detect viruses at relevant concentrations for diagnostic applications. Hence, a novel assay was developed that allows simultaneous concentration and detection of viruses using a microfluidic chip with an integrated nanoporous membrane. The size-exclusion properties of the in situ polymerized polyacrylamide membrane are exploited to simultaneously concentrate viral particles and separate the virus/fluorescent antibody complex from the unbound antibody. The assay is performed in two simple steps-addition of fluorescently labeled antibodies to the sample, followed by concentration of antibody-virus complexes on a porous membrane. Excess antibodies are removed by electrophoresis through the membrane and the complex is then detected downstream of the membrane. This new assay detected inactivated swine influenza virus at a concentration four times lower than that of the open-channel electrophoresis assay. The total assay time, including device regeneration, is six minutes and requires <50 μl of sample. The filtration effect of the polymer membrane eliminates the need for washing, commonly required with surface-based immunoassays, increasing the speed of the assay. This assay is intended to form the core of a portable device for the diagnosis of high-consequence animal pathogens such as foot-and-mouth disease. The electrophoretic immunoassay format is rapid and simple while providing the necessary sensitivity for diagnosis of the illness state. This would allow the development of a portable, cost-effective, on-site diagnostic system for rapid screening of large populations of livestock, including sheep, pigs, cattle, and potentially birds. © The Royal Society of Chemistry.

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The Bio-Restoration of Major Transportation Facilities Domestic Demonstration and Application Program (DDAP) is a designed to accelerate the restoration of transportation nodes following an attack with a biological warfare agent. This report documents the technology development work done at SNL for this DDAP, which include development of the BROOM tool, an investigation of surface sample collection efficiency, and a flow cytometry study of chlorine dioxide effects on Bacillus anthracis spore viability.

In February of 2005, a joint exercise involving Sandia National Laboratories (SNL) and the National Institute for Occupational Safety and Health (NIOSH) was conducted in Albuquerque, NM. The SNL participants included the team developing the Building Restoration Operations and Optimization Model (BROOM), a software product developed to expedite sampling and data management activities applicable to facility restoration following a biological contamination event. Integrated data-collection, data-management, and visualization software improve the efficiency of cleanup, minimize facility downtime, and provide a transparent basis for reopening. The exercise was held at an SNL facility, the Coronado Club, a now-closed social club for Sandia employees located on Kirtland Air Force Base. Both NIOSH and SNL had specific objectives for the exercise, and all objectives were met.

The natural gas industry seeks inexpensive sensors and instrumentation to rapidly measure gas heating value in widely distributed locations. For gas pipelines, this will improve gas quality during transfer and blending, and will expedite accurate financial accounting. Industrial endusers will benefit through continuous feedback of physical gas properties to improve combustion efficiency during use. To meet this need, Sandia has developed a natural gas heating value monitoring instrument using existing and modified microfabricated components. The instrument consists of a silicon micro-fabricated gas chromatography column in conjunction with a catalytic micro-calorimeter sensor. A reference thermal conductivity sensor provides diagnostics and surety. This combination allows for continuous calorimetric determination with a 1 minute analysis time and 1.5 minute cycle time using air as a carrier gas. This system will find application at remote natural gas mining stations, pipeline switching and metering stations, turbine generators, and other industrial user sites. Microfabrication techniques will allow the analytical components to be manufactured in production quantities at a low per-unit cost.

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This slide presentation outlines information on a technology acquisition strategy for the security of water distribution networks. The Department of Homeland Security (DHS) has tasked a multi-laboratory team to evaluate current and future needs to protect the nation's water distribution infrastructure by supporting an objective evaluation of current and new technologies. The primary deliverables from this Operational Technology Demonstration (OTD) are the following: establishment of an advisory board for review and approval of testing protocols, technology acquisition processes and recommendations for technology test and evaluation in laboratory and field settings; development of a technology acquisition process; creation of laboratory and field testing and evaluation capability; and, testing of candidate technologies for insertion into a water early warning system. The initial phase of this study involves the development of two separate but complementary strategies to be reviewed by the advisory board: a technology acquisition strategy; and, a technology evaluation strategy. Lawrence Livermore National Laboratory and Sandia National Laboratories are tasked with the first strategy, while Los Alamos, Pacific Northwest, and Oak Ridge National Laboratories are tasked with the second strategy. The first goal of the acquisition strategy is the development of a technology survey process that includes a review of current test programs and development of a method to solicit and select existing and emerging sensor technologies for evaluation and testing. The second goal is to implement the acquisition strategy to provide a set of recommendations for candidate technologies for laboratory and field testing.

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This work describes the design, simulation, fabrication and characterization of a microfabricated thermal conductivity detector to be used as an extension of the {micro}ChemLab{trademark}. The device geometry was optimized by simulating the heat transfer in the device, utilizing a boundary element algorithm. In particular it is shown that within microfabrication constraints, a micro-TCD optimized for sensitivity can be readily calculated. Two flow patterns were proposed and were subsequently fabricated into nine-promising geometries. The microfabricated detector consists of a slender metal film, supported by a suspended thin dielectric film over a pyramidal or trapezoidal silicon channel. It was demonstrated that the perpendicular flow, where the gas directly impinges on the membrane, creates a device that is 3 times more sensitive than the parallel flow, where the gas passed over the membrane. This resulted in validation of the functionality of a microfabricated TCD as a trace-level detector, utilizing low power. the detector shows a consistent linear response to concentration and they are easily able to detect 100-ppm levels of CO in He. Comparison of noise levels for this analysis indicates that sub part per million (ppm) levels are achievable with the selection of the right set of conditions for the detector to operate under. This detector was originally proposed as part of a high-speed detection system for the petrochemical gas industry. This system was to be utilized as a process monitor to detect reactor ''upset'' conditions before a run away condition could occur (faster than current full-scale monitoring systems were able to achieve). Further outlining of requirements indicated that the detection levels likely achievable with a TCD detector would not be sufficient to meet the process condition needs. Therefore the designed and fabricated detector was integrated into a detection system to showcase some technologies that could further the development of components for the current gas phase {micro}ChemLab as well as future modifications for process monitoring work such as: pressurized connections, gas sampling procedures, and packed columns. Component integration of a microfabricated planar pre-concentrator, gas-chromatograph column and TCD in the separation/detection of hydrocarbons, such as benzene, toluene and xylene (BTX) was also demonstrated with this system.