Rapid viability polymerase chain reaction method for detection of Francisella tularensis

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. Due to slow growth on even selective culture media, detection of viable F. tularensis from environmental and drinking water samples, usually takes >3 days. Therefore, a rapid viability polymerase chain reaction (RV-PCR) method was developed to detect and identify viable F. tularensis cells in environmental samples. The method uses a change in PCR response during high throughput (48-well) sample incubation in Brain Heart Infusion/Vitox/Fildes/Histidine growth medium to detect viable F. tularensis presence, which is at least two times faster than the current plate culture-based method. Using the method, 10¹ to 10² live F. tularensis cells were detected in simulated complex sample matrices containing chemical and biological interferences.