Nguyen C, Makkar R, Sharp N J, Page M A, Molineux I J, Schofield D A
Guild BioSciences, Charleston, SC, USA.
Construction Engineering and Research Laboratory, U.S. Army Corps of Engineers, Engineer Research and Development Center, Champaign, IL, USA.
J Appl Microbiol. 2017 Nov;123(5):1184-1193. doi: 10.1111/jam.13569. Epub 2017 Sep 21.
We investigated the ability of a temperate Bacillus anthracis reporter phage (Wβ::luxAB-2), which transduces bioluminescence to infected cells, to detect viable spores from deliberately contaminated environmental water samples.
Environmental water was inoculated with spores and assayed with Wβ::luxAB-2. Bioluminescent signals directly correlated with input phage and spore concentrations. A limit of detection of 10 and 10 CFU per ml within 8 h was achieved from pond and lake water, respectively. Detection was greatly simplified by minimizing sample processing steps without spore extraction. The complex endogenous microbial flora and salt content of brackish water challenged the assay, extending the detection time to 12 h for a sensitivity of 10 CFU per ml. Phage-mediated bioluminescence was strictly dependent on bacterial physiology, being significantly reduced in mid/late log phase cells. This was shown to be due to an inability of the phage to adsorb.
The reporter phage Wβ::luxAB-2 displays potential for simplified detection of viable spores from contaminated water samples within 12 h.
A deliberate aerosol release of spores could lead to widespread contamination, leaving large areas uninhabitable until remediation. An essential requirement of this restoration process is the development of simplified detection assays in different environmental matrices.
我们研究了一种温和型炭疽芽孢杆菌报告噬菌体（Wβ::luxAB-2）将生物发光转导至受感染细胞的能力，以检测故意污染的环境水样中的活孢子。
将孢子接种到环境水样中，并用Wβ::luxAB-2进行检测。生物发光信号与输入的噬菌体和孢子浓度直接相关。分别从池塘水和湖水中在8小时内实现了每毫升10和10 CFU的检测限。通过尽量减少无孢子提取的样品处理步骤，检测得到了极大简化。微咸水复杂的内源微生物群落和盐分含量对检测造成了挑战，将检测时间延长至12小时，灵敏度为每毫升10 CFU。噬菌体介导的生物发光严格依赖于细菌生理学，在对数中期/后期细胞中显著降低。这表明是由于噬菌体无法吸附。
报告噬菌体Wβ::luxAB-2具有在12小时内简化检测污染水样中活孢子的潜力。
故意气溶胶释放孢子可能导致广泛污染，在进行修复之前大片区域无法居住。这一恢复过程的一项基本要求是开发在不同环境基质中的简化检测方法。
