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Method Development for Detection of Biothreat Agents from Environmental Samples
Oral Presentation
Prepared by T. Nichols1, S. Shah2, H. Ernst3
1 - USEPA NHSRC, 1200 Pennsylvania NW, Washington, DC, 20460
2 - USEPA, 1200 Pennsylvania NW, Washington, DC, 20460
3 - USEPA, 26 Martin Luther King, Cincinnati, OH, 45268
Contact Information: nichols.tonya@epa.gov; 202-564-2338
ABSTRACT
In the event of an indoor, outdoor, or water system contamination with pathogenic microorganisms, it is necessary to have sampling and analytical methods that measure the concentration of the biological agent of concern in the environmental samples taken from the various environmental matrices (water, surfaces, and air). To address this need, the Homeland Security Research Program (HSRP) conducts research to select existing methods for immediate use or further refinement of the method to better fit the response community’s requirements for sensitive, higher-throughput methods in a timely and cost-effective manner for risk management decisions. Our method development efforts are in direct support of the EPA’s Environmental Response Laboratory Network (ERLN). The biological detection methods research includes both biological select and non-select agents with Bacillus anthracis being the highest priority. Standard Analytical Procedures (SAPs) have been developed for several biothreat agents. Few of the SAPs including those for Escherichia coli O157:H7, Vibrio cholerae, Salmonella Typhi, and Non-Typhoidal Salmonella species in water samples have been verified in a single laboratory set up. On the select agents, our efforts have been focused on combining the traditional culture method with high-throughput molecular analysis using the polymerase chain reaction (PCR) for the detection of B. anthracis spores. The Rapid Viability PCR (RV-PCR) method has been developed and optimized for air filters, wipes, water, vacuum filters, vacuum socks, and sponge-stick sampling tools. Single laboratory verification of the method demonstrated a limit of detection (LOD) at the level of 10 CFU/sample for air filters, wipes, and water samples. The method was field-tested using B. anthracis surrogate samples from a facility decontamination exercise. In a split-sample analysis, the RV-PCR method was 97.8% in agreement with traditional culture-based method. In summary, our research aims to provide rapid, high-throughput, highly sensitive, specific, time-efficient, and cost-effective methods for the detection of viable biological contaminants. This presentation will describe method development and testing for biothreat agents and the applicability to recovery from contamination incidents.
Oral Presentation
Prepared by T. Nichols1, S. Shah2, H. Ernst3
1 - USEPA NHSRC, 1200 Pennsylvania NW, Washington, DC, 20460
2 - USEPA, 1200 Pennsylvania NW, Washington, DC, 20460
3 - USEPA, 26 Martin Luther King, Cincinnati, OH, 45268
Contact Information: nichols.tonya@epa.gov; 202-564-2338
ABSTRACT
In the event of an indoor, outdoor, or water system contamination with pathogenic microorganisms, it is necessary to have sampling and analytical methods that measure the concentration of the biological agent of concern in the environmental samples taken from the various environmental matrices (water, surfaces, and air). To address this need, the Homeland Security Research Program (HSRP) conducts research to select existing methods for immediate use or further refinement of the method to better fit the response community’s requirements for sensitive, higher-throughput methods in a timely and cost-effective manner for risk management decisions. Our method development efforts are in direct support of the EPA’s Environmental Response Laboratory Network (ERLN). The biological detection methods research includes both biological select and non-select agents with Bacillus anthracis being the highest priority. Standard Analytical Procedures (SAPs) have been developed for several biothreat agents. Few of the SAPs including those for Escherichia coli O157:H7, Vibrio cholerae, Salmonella Typhi, and Non-Typhoidal Salmonella species in water samples have been verified in a single laboratory set up. On the select agents, our efforts have been focused on combining the traditional culture method with high-throughput molecular analysis using the polymerase chain reaction (PCR) for the detection of B. anthracis spores. The Rapid Viability PCR (RV-PCR) method has been developed and optimized for air filters, wipes, water, vacuum filters, vacuum socks, and sponge-stick sampling tools. Single laboratory verification of the method demonstrated a limit of detection (LOD) at the level of 10 CFU/sample for air filters, wipes, and water samples. The method was field-tested using B. anthracis surrogate samples from a facility decontamination exercise. In a split-sample analysis, the RV-PCR method was 97.8% in agreement with traditional culture-based method. In summary, our research aims to provide rapid, high-throughput, highly sensitive, specific, time-efficient, and cost-effective methods for the detection of viable biological contaminants. This presentation will describe method development and testing for biothreat agents and the applicability to recovery from contamination incidents.