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Development of a Rapid Viability Polymerase Chain Reaction Method for Detection of Yersinia pestis in Water
Kane, S., S. Shah, AND T. Alfaro. Development of a Rapid Viability Polymerase Chain Reaction Method for Detection of Yersinia pestis in Water. JOURNAL OF MICROBIOLOGICAL METHODS. Elsevier Science Ltd, New York, NY, 162:21-27, (2019).
To protect the human health and the environment, the EPA decision makers will need timely and reliable water sample analysis results during the response to and recovery from a plague incident resulting in contamination of the water infrastructure regardless of whether the contamination is a result of a natural outbreak, laboratory accident, or a criminal or terrorist incident. This report presents the research effort focused on the development and optimization of the Rapid Viability Polymerase Chain Reaction (RV-PCR) method for detection of Yersinia pestis (the bacteria that cause plague) in water samples. The RV-PCR method rapidly determines the presence or absence of live Y. pestis in a sample. The method can be less labor-, space-, and time- intensive than the traditional microbiological culture methods and will address the need for rapid sample analysis. The method will help enhance the capability and capacity for rapid, reliable, and high throughput sample analysis for the Water Laboratory Alliance (WLA), a network of laboratories established by the EPA Office of Water.
Due to the occurrence of natural plague outbreaks and its historical usage as a biological weapon, Yersinia pestis is considered one of the high-priority biological threat agents. It can remain viable in certain environments including water for more than 100 days. The Rapid Viability Polymerase Chain Reaction (RV-PCR) method combines shorter sample incubation in liquid culture (compared to plate culture) with real-time PCR analysis before and after incubation and uses the change in real-time PCR response to specifically detect low concentrations of viable Y. pestis.
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DEVELOPMENT OF A RAPID VIABILITY POLYMERASE CHAIN REACTION METHOD FOR DETECTION OF YERSINIA PESTIS Exit