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Comprehensive Laboratory Evaluation of a Lateral Flow Assay for the Detection of Yersinia pestis
Kristin W Prentice1, Lindsay DePalma1, Jason G Ramage1
1Kristin W. Prentice, MS, is an Associate, Booz Allen Hamilton, Rockville, MD. Lindsay DePalma, MS, is a Staff Life Scientist, Booz Allen Hamilton, McLean, VA. Jason G. Ramage, MS, MBA, PMP, is Assistant Vice Chancellor for Research and Innovation and Director of Research Compliance, University of Arkansas, Fayetteville, AR. Jawad Sarwar, MS, is a Senior Research Scientist, and Nishanth Parameswaran and Mrinmayi Joshi, MS, are Research Scientists; all at Omni Array Biotechnology, Rockville, MD. Jeannine Petersen, PhD, Brook Yockey, and John Young are Microbiologists; all with DHHS/CDC/OID/NCEZID/DVBD/BDB, Fort Collins, CO. Nagarajan Thirunavvukarasu, PhD, is an ORISE Fellow; Christine A. Pillai and Gowri Manickam, PhD, are ORISE Fellow Research Scientists; and Shashi K. Sharma, PhD, is a Research Microbiologist; all at the FDA Center for Food Safety and Applied Nutrition, Molecular Methods Development Branch, Division of Microbiology, Office of Regulatory Science, College Park, MD. Ajay Singh, PhD, is a Research Scientist, Laulima Government Solutions, Contractor Support to USAMRICD, Neurobiological Toxicology Branch, Analytical Toxicology Division, Aberdeen Proving Ground, MD. Carol Chapman, MS, is a Microbiologist, Geneva Foundation, Contractor Support to the Naval Medical Research Center, Silver Spring, MD. Julie R. Avila, MS, is a Scientific Associate, Lawrence Livermore National Laboratory, Biosciences and Biotechnology Division, Livermore, CA. Stephen A. Morse, MSPH, PhD, is a Senior Advisor, CDC Division of Select Agents and Toxins, and is currently with IHRC, Inc., Atlanta, GA. Kodumudi Venkat Venkateswaran, PhD, is Chief Scientist, Tetracore, Inc., Rockville, MD. Kevin Anderson, PhD, and David R. Hodge, PhD, are Program Managers, Science and Technology Directorate, US Department of Homeland Security, Washington, DC. Segaran P. Pillai, PhD, is Director, Office of Laboratory Science and Safety, FDA Office of the Commissioner, Silver Spring, MD.
Abstract:
We conducted a comprehensive, multiphase laboratory evaluation of the Plague BioThreat Alert® (BTA) test, a lateral flow immunoassay (LFA), for the rapid detection of Yersinia pestis. The study was conducted in 7 phases at 2 sites to assess the performance of the LFA. The limit of detection (LOD) was determined using both a virulent and avirulent strain of Y. pestis, CO99-3015 (105 CFU/ml) and A1122 (104 CFU/ml), respectively. In the other phases, 18 Y. pestis strains, 20 phylogenetic near-neighbor strains, 61 environmental background microorganisms, 26 white powders, and a pooled aerosol sample were also tested. A total of 1,110 LFA test results were obtained, and their analysis indicates that this LFA had a sensitivity of 97.65% and specificity of 96.57%. These performance data are important for accurate interpretation of qualitative results arising from testing suspicious white powders and aerosol samples in the field. Any positive specimen in this assay is considered presumptive positive and should be referred to the Centers for Disease Control and Prevention Laboratory Response Network for additional testing, confirmation, and characterization for an appropriate public health response.
Insights
The Plague BioThreat Alert (BTA) test, a lateral flow immunoassay, accurately detects Yersinia pestis. This rapid test shows high sensitivity and specificity for identifying potential biothreat agents.
Area of Science:
- Microbiology
- Immunology
- Public Health
Background:
- Yersinia pestis is a dangerous bacterium that causes plague.
- Rapid detection of Yersinia pestis is crucial for public health response.
- Lateral flow immunoassays (LFAs) offer potential for rapid diagnostics.
Purpose of the Study:
- To evaluate the performance of the Plague BioThreat Alert (BTA) test, a lateral flow immunoassay (LFA).
- To assess the sensitivity and specificity of the BTA test for detecting Yersinia pestis.
- To determine the utility of the BTA test in identifying Yersinia pestis in environmental samples.
Main Methods:
- A comprehensive, multiphase laboratory evaluation of the BTA test was conducted.
- The limit of detection (LOD) was determined using virulent and avirulent Yersinia pestis strains.
- The assay was tested against various Yersinia pestis strains, near-neighbor organisms, environmental microorganisms, and white powders.
Main Results:
- A total of 1,110 test results were analyzed.
- The BTA test demonstrated a sensitivity of 97.65% and a specificity of 96.57%.
- Performance data were generated for detecting Yersinia pestis in white powders and aerosol samples.
Conclusions:
- The Plague BioThreat Alert (BTA) test is a reliable tool for the rapid detection of Yersinia pestis.
- The LFA exhibits high sensitivity and specificity, making it valuable for field testing.
- Positive results are presumptive and require confirmation by reference laboratories for public health actions.
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