Takeuchi Ippei, Nasukawa Tadahiro, Sugimoto Ryosuke, Takemura-Uchiyama Iyo, Murakami Hironobu, Uchiyama Jumpei
School of Veterinary Medicine, Azabu University, Kanagawa, Japan.
School of Veterinary Medicine, Azabu University, Kanagawa, Japan.
Virus Res. 2021 Jun;298:198406. doi: 10.1016/j.virusres.2021.198406. Epub 2021 Mar 30.
To introduce phage therapy against multidrug-resistant Staphylococcus aureus in Western medicine, the establishment of phage manufacturing, particularly phage propagation, is indispensable. For the propagation of S. aureus phages, knowledge of the effects of phage types, process parameters, and analytical methodologies should be investigated. In this study, S. aureus phage propagations were studied in a flask with a new class of design of experiments, definitive screening design, using S. aureus phages S13' and S25-3 in different taxonomies. Four process parameters, namely, multiplicity of infection, bacterial density at infection, time of harvest, and temperature, were evaluated with the regression models based on the phage concentration data measured using plaque assay and quantitative polymerase chain reaction. As a result, phage propagations measured using plaque assay and quantitative polymerase chain reaction were overall similar to each other in the case of phage S13', while they differed in the case of phage S25-3. These results suggest that the propagation processes need to be developed according to phage type, and the choice of methodologies for phage concentration measurements should be carefully considered.
在西医中引入针对耐多药金黄色葡萄球菌的噬菌体疗法时，建立噬菌体生产，尤其是噬菌体增殖，是必不可少的。对于金黄色葡萄球菌噬菌体的增殖，应研究噬菌体类型、工艺参数和分析方法的影响。在本研究中，使用新型实验设计——确定性筛选设计，在烧瓶中研究了不同分类学中的金黄色葡萄球菌噬菌体S13'和S25-3的增殖情况。基于噬菌斑测定和定量聚合酶链反应测得的噬菌体浓度数据，利用回归模型评估了四个工艺参数，即感染复数、感染时的细菌密度、收获时间和温度。结果，在噬菌体S13'的情况下，使用噬菌斑测定和定量聚合酶链反应测得的噬菌体增殖总体上彼此相似，而在噬菌体S25-3的情况下则有所不同。这些结果表明，需要根据噬菌体类型来开发增殖工艺，并且应仔细考虑噬菌体浓度测量方法的选择。
