一种改进的抗呼吸道合胞病毒药物筛选方法  

Improvements in methods for screening potential antiviral agents agains t respiratory syncytial virus

作　　者：方学平[1] 张振亚[1] 曾瑞红[1] 刁志花[1] 梅兴国[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《军事医学科学院院刊》2005年第4期352-354,共3页Bulletin of the Academy of Military Medical Sciences

摘　　要：目的:在细胞病变抑制实验的基础上,对观察指标和结果评价的方法进行改进,建立一种快速、可靠的筛选抗呼吸道合胞病毒药物的方法.方法:通过观察计数接种病毒后出现的病变数目,确定改进实验中最佳的细胞接种密度和结果判定时间,并推算药物的半数有效浓度EC50.比较空斑减数实验和改进方法测定利巴韦林(病毒唑)的EC50的差异.结果:改进方法和空斑减数实验测得的利巴韦林的EC50平均值没有显著性差异(P＞0.05).最佳实验条件是:每孔接种Hep-2细胞(1.5～2.0)×104个为宜,24～36 h接种病毒,第4～5天可判读最后结果.结论:该方法具有准确可靠、简单和快速的特点,可以应用于抗呼吸道合胞病毒药物的筛选.Objective：To establish a rapid and reliable method for screening potential antiviral agents against respiratory syncytial virus based on the cytopathogenic effect inhibition assay. Methods： Hep-2 cells of different inoculated density were infected by RSV and the numbers of lesions of cytopathic effect（CPE） were counted everyday. The appropriate experiment conditions were determinated through evaluation of cell growth and the development of CPE. The antiviral effect of ribavirin was tested both by the plaque reduction assay and the improved method, and the results were analyzed with ANOVA by the software of SAS. Results： The average EC50 had no significant difference with the plaque reduction assay （ P 〉 0.05 ）. The appropriate experimental conditions were as follows ： Hep-2 cells were inoculated in 96-well plastic plates about （ 1.5 - 2.0） × 10^4 per well, virus suspension containing an appropriate concentrations of the test compounds were inoculated after 24 - 36 h, CPE was evaluated after 4 - 5 days of incubations. Conclusion ： The improved method for screening antiviral agents against respiratory syncytial virus is reliable, simple and rapid.

关 键 词：呼吸道合胞病毒 抗病毒药 药物筛选 

分 类 号：R965.1[医药卫生—药理学]