Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, Innovation Academy of South China Sea Ecology and Environmental Engineering, South China Sea Institute of Oceanology, Chinese Academy of Sciences, No.1119, Haibin Road, Nansha District, Guangzhou, 511458, China.
Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou), No.1119, Haibin Road, Nansha District, Guangzhou, 511458, China.
Environ Microbiol Rep. 2021 Oct;13(5):728-734. doi: 10.1111/1758-2229.12991. Epub 2021 Jul 10.
Phage contamination is a common complication for the fermentation and pharmaceutical industries. The risk of bacteriophage contamination in laboratory processes increases with multiple rounds of genetic manipulation such as deletion and complementation. The contamination of temperate phages does not lead to immediate host cell lysis but could become a serious issue when the lytic cycle is activated under specific conditions. Our objective was to develop a quick and reliable detection method for checking possible temperate phage contamination. Here, using motility plates, we found that when the strain carries a newly acquired temperate phage, its presence can be easily detected by the formation of a clear 'lysis zone' when swimming against the original strain on the same swimming plates. Compared to the traditional double agar layer method and genomic sequencing-based methods, the duration of the motility-based assay is shorter and the procedure is simplified. More importantly, for the bacterial strains that already contain active prophages, this method can still easily detect the newly acquired phages without tedious phage identification procedure. These features make this method highly applicable to laboratory and industrial processes.
噬菌体污染是发酵和制药行业的常见并发症。在实验室过程中，随着诸如缺失和互补等多次遗传操作，噬菌体污染的风险会增加。温和噬菌体的污染不会导致宿主细胞立即裂解，但当在特定条件下激活裂解周期时，可能会成为一个严重的问题。我们的目标是开发一种快速可靠的检测方法，以检查可能存在的温和噬菌体污染。在这里，我们使用游动平板发现，当菌株携带新获得的温和噬菌体时，当它在同一游动平板上与原始菌株反向游动时，形成清晰的“裂解区”，其存在可以很容易地被检测到。与传统的双层琼脂层方法和基于基因组测序的方法相比，基于运动的测定法耗时更短，程序更简化。更重要的是，对于已经含有活性原噬菌体的细菌菌株，该方法仍然可以轻松检测到新获得的噬菌体，而无需繁琐的噬菌体鉴定程序。这些特点使该方法非常适用于实验室和工业过程。
