Lawrence Livermore National Laboratory, Livermore, CA, USA.
National Homeland Security Research Center, Office of Research and Development, U.S. Environmental Protection Agency, Washington, DC, USA.
J Microbiol Methods. 2019 Nov;166:105738. doi: 10.1016/j.mimet.2019.105738. Epub 2019 Oct 15.
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, 10 to 10 live F. tularensis cells were detected in simulated complex sample matrices containing chemical and biological interferences.
弗朗西斯菌，会引起致命的土拉菌病，由于其感染剂量低、报告的环境持久性以及能够通过多种暴露途径传播给人类，被认为是生物恐怖主义和生物战的一种有吸引力的制剂。由于即使在选择性培养基上生长也很缓慢，因此从环境和饮用水样本中检测到存活的弗朗西斯菌通常需要>3 天。因此，开发了一种快速存活聚合酶链反应（RV-PCR）方法来检测和鉴定环境样本中的存活弗朗西斯菌细胞。该方法使用在脑心浸液/ Vitox/ Fildes/组氨酸生长培养基中进行高通量（48 孔）样本孵育过程中 PCR 反应的变化来检测存活的弗朗西斯菌的存在，其速度至少比当前基于平板培养的方法快两倍。使用该方法，在含有化学和生物干扰物的模拟复杂样本基质中，可以检测到 10 到 10 个活的弗朗西斯菌细胞。
