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RAPID VIABILITY POLYMERASE CHAIN REACTION METHOD FOR DETECTION OF FRANCISELLA TULARENSIS
Kane, S., S. Shah, AND T. Alfaro. RAPID VIABILITY POLYMERASE CHAIN REACTION METHOD FOR DETECTION OF FRANCISELLA TULARENSIS. JOURNAL OF MICROBIOLOGICAL METHODS. Elsevier Science Ltd, New York, NY, 166:105738, (2019). https://doi.org/10.1016/j.mimet.2019.105738
Francisella tularensis, the pathogen that causes tularemia in humans and animals, could be introduced into water infrastructure due to a natural outbreak, laboratory accident, or intentional contamination. This pathogen survives for many days in the environment and water. It is a very slow growing pathogen and can take several days to detect using the traditional microbiological culture methods. Therefore, a need for a rapid and high-throughput analytical method was identified and EPA addressed by developing and optimizing a Rapid Viability PCR method to relatively rapidly detect F. tularensis. Using this method, results can be obtained in 36 hours as compared to several days by the traditional microbiological culture methods. Also, with this method, high-throughput sample analysis can be performed using a 48-well culture plate which requires very low volume of growth medium resulting in additional advantage of significantly low laboratory waste. This method is developed for use by the EPA’s Environmental Response Laboratory Network (ERLN) that includes the Water Laboratory Alliance (WLA) network managed by the EPA Office of Water. It can also be useful to other Federal Agencies’ laboratory networks for a national level need.
Francisella tularensis, which causes potentially fatal tularemia, has been considered an attractive agent of bioterrorism and biological warfare due to its low infectious dose, reported environmental persistence, and ability to be transmitted to humans via multiple exposure routes. EPA developed a high-throughput Rapid Viability-PCR protocol to relatively rapidly detect Francisella tularensis. This study contains summarized data from multiple experimental results to show the method performance in various experimental conditions mimicking a real-world interference material.
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