4.4 Article

Rapid detection of temperate bacteriophage using a simple motility assay

期刊

ENVIRONMENTAL MICROBIOLOGY REPORTS
卷 13, 期 5, 页码 728-734

出版社

WILEY
DOI: 10.1111/1758-2229.12991

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资金

  1. National Science Foundation of China [31625001, 31970037, 91951203]
  2. Guangdong Major Project of Basic and Applied Basic Research [2019B030302004]
  3. Guangdong Local Innovation Team Program [2019BT02Y262]
  4. Innovation Academy of South China Sea Ecology and Environmental Engineering, the Chinese Academy of Sciences [ISEE2018PY01]
  5. Key Special Project for Introduced Talents Team of Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou) [GML2019ZD0407]

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The text discusses the common complication of phage contamination in the fermentation and pharmaceutical industries, and introduces a quick and reliable detection method through motility-based assay for possible temperate phage contamination. Compared to traditional methods and genomic sequencing-based methods, this approach has a shorter duration and a simplified procedure.
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.

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