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A modified MS2 bacteriophage plaque reduction assay for the rapid screening of antiviral plant extracts

机译:用于快速筛选抗病毒植物提取物的改良MS2噬菌体噬斑减少法

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Introduction:Traditional methods of screening plant extracts and purified components for antiviral activity require up to a week to perform, prompting the need to develop more rapid quantitative methods to measure the ability of plant based preparations to block viral replication. We describe an adaption of an MS2 plaque reduction assay for use in S. aureus.Results:MS2 bacteriophage was capable of infecting and replicating in B. cereus, S. aureus and F + E. coli but not F- E. coli. Indeed, both B. cereus and S. aureus were more sensitive to MS2 induced lysis than F+ E. coli. When MS2 bacteriophage was mixed with Camellia sinensis extract (1 mg/ml), Scaevola spinescens extract (1 mg/ml) or Aloe barbadensis juice and the mixtures inoculated into S. aureus, the formation of plaques was reduced to 8.9 ± 3.8%, 5.4 ± 2.4% and 72.7 ± 20.9% of the untreated MS2 control values respectively.Conclusions:The ability of the MS2 plaque reduction assay to detect antiviral activity in these known antiviral plant preparations indicates its suitability as an antiviral screening tool. An advantage of this assay compared with traditionally used cytopathic effect reduction assays and replicon based assays is the more rapid acquisition of results. Antiviral activity was detected within 24 h of the start of testing. The MS2 assay is also inexpensive and non-pathogenic to humans making it ideal for initial screening studies or as a simulant for pathogenic viruses.
机译:简介:筛选植物提取物和纯化成分的抗病毒活性的传统方法需要长达一周的时间才能完成,这提示需要开发更快速的定量方法来测量基于植物的制剂阻断病毒复制的能力。结果:MS2噬菌体能够在蜡样芽孢杆菌,金黄色葡萄球菌和F +大肠杆菌中感染和复制,但不能在F-大肠杆菌中感染和复制。实际上,蜡状芽孢杆菌和金黄色葡萄球菌对MS2诱导的裂解均比F +大肠杆菌更敏感。当将MS2噬菌体与山茶提取物(1 mg / ml),棘孢茅(Scaevola spinescens)提取物(1 mg / ml)或库拉索芦荟汁混合,并将混合物接种到金黄色葡萄球菌中时,噬菌斑的形成减少到8.9±3.8%结论:在这些已知的抗病毒植物制剂中,MS2噬斑减少检测法检测抗病毒活性的能力表明其作为抗病毒筛选工具的适用性。分别为未经处理的MS2对照值的5.4±2.4%和72.7±20.9%。与传统使用的细胞病变效应降低测定法和基于复制子的测定法相比,该测定法的优点是更快地获得结果。在测试开始后的24小时内检测到抗病毒活性。 MS2测定法也很便宜,并且对人类无致病性,因此非常适合进行初始筛选研究或用作病原病毒的模拟物。

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