Liu Weili, Li Chao, Qiu Zhi-Gang, Jin Min, Wang Jing-Feng, Yang Dong, Xiao Zhong-Hai, Yuan Zhao-Kang, Li Jun-Wen, Xu Qun-Ying, Shen Zhi-Qiang
Tianjin Institute of Health and Environmental Medicine, Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Tianjin 300050, China.
School of Public Health, Nanchang University, Jiangxi 330006, China.
J Microbiol Methods. 2017 Aug;139:143-149. doi: 10.1016/j.mimet.2017.05.019. Epub 2017 May 31.
Bacteriophages are widely used to the treatment of drug-resistant bacteria and the improvement of food safety through bacterial lysis. However, the limited investigations on bacteriophage restrict their further application. In this study, a novel and highly efficient method was developed for isolating bacteriophage from water based on the electropositive silica gel particles (ESPs) method. To optimize the ESPs method, we evaluated the eluent type, flow rate, pH, temperature, and inoculation concentration of bacteriophage using bacteriophage f2. The quantitative detection reported that the recovery of the ESPs method reached over 90%. The qualitative detection demonstrated that the ESPs method effectively isolated 70% of extremely low-concentration bacteriophage (10 PFU/100L). Based on the host bacteria composed of 33 standard strains and 10 isolated strains, the bacteriophages in 18 water samples collected from the three sites in the Tianjin Haihe River Basin were isolated by the ESPs and traditional methods. Results showed that the ESPs method was significantly superior to the traditional method. The ESPs method isolated 32 strains of bacteriophage, whereas the traditional method isolated 15 strains. The sample isolation efficiency and bacteriophage isolation efficiency of the ESPs method were 3.28 and 2.13 times higher than those of the traditional method. The developed ESPs method was characterized by high isolation efficiency, efficient handling of large water sample size and low requirement on water quality.
噬菌体被广泛应用于耐药菌的治疗以及通过细菌裂解来提高食品安全。然而，对噬菌体的有限研究限制了它们的进一步应用。在本研究中，基于正电硅胶颗粒（ESPs）方法开发了一种从水中分离噬菌体的新型高效方法。为了优化ESPs方法，我们使用噬菌体f2评估了洗脱液类型、流速、pH值、温度和噬菌体接种浓度。定量检测表明，ESPs方法的回收率达到90%以上。定性检测表明，ESPs方法能有效分离70%的极低浓度噬菌体（10 PFU/100L）。基于由33株标准菌株和10株分离菌株组成的宿主细菌，采用ESPs方法和传统方法从天津海河流域三个地点采集的18份水样中分离噬菌体。结果表明，ESPs方法明显优于传统方法。ESPs方法分离出32株噬菌体，而传统方法分离出15株。ESPs方法的样品分离效率和噬菌体分离效率分别比传统方法高3.28倍和2.13倍。所开发的ESPs方法具有分离效率高、能有效处理大体积水样且对水质要求低的特点。
