Cock Ian, Kalt F R
Biomolecular and Physical Sciences, Nathan Campus, Griffith University, 170 Kessels Road, Nathan, Queensland 4111, Australia.
Pharmacognosy Res. 2010 Jul;2(4):221-8. doi: 10.4103/0974-8490.69108.
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.
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.
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.
传统的筛选植物提取物和纯化成分抗病毒活性的方法需要长达一周的时间来完成，这促使人们需要开发更快速的定量方法来测量植物制剂阻断病毒复制的能力。我们描述了一种适用于金黄色葡萄球菌的MS2噬菌斑减少试验的改良方法。
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。事实上，蜡样芽孢杆菌和金黄色葡萄球菌对MS2诱导的裂解比F+大肠杆菌更敏感。当MS2噬菌体与1毫克/毫升的中华茶提取物、1毫克/毫升的刺草海桐提取物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，噬菌斑的形成分别减少到未处理的MS2对照值的8.9±3.8%、5.4±2.4%和72.7±20.9%。
MS2噬菌斑减少试验检测这些已知抗病毒植物制剂中抗病毒活性的能力表明其适合作为一种抗病毒筛选工具。与传统使用的细胞病变效应减少试验和基于复制子的试验相比，该试验的一个优点是能更快地获得结果。在测试开始后24小时内检测到了抗病毒活性。MS2试验成本低廉且对人类无致病性，使其成为初始筛选研究或作为致病病毒模拟物的理想选择。
