细胞荧光转化灶单位实验体外滴定狂犬病病毒毒力方法的优化

Optimization of Method of Titration Rabies Virus with Fluorescence Focus Units Assay

目的用细胞荧光转化灶单位实验[Fluorescence Focus Units(FFU)Assay]替代小鼠50%致死量(50% Lethal Does,LD50)法检测狂犬病病毒毒力,优化、改进现有的检测方法 ,从而降低检测成本。方法将狂犬病病毒固定毒株3倍系列稀释后感染乳金黄地鼠肾细胞,通过荧光标记的抗狂犬病病毒核蛋白单克隆抗体染色检测病毒感染的荧光灶,同时将病毒颅内接种10～12g昆明小鼠,通过观察记录小鼠发病死亡数,计算引起半数小鼠死亡的病毒剂量即LD50。结果 FFU和LD50存在一定的数量关系,且成正相关,相关系数为0.99。lgFFU值与LD50值之间有统计学意义,重复性好。结论 FFU实验体外滴定狂犬病病毒毒力方法可取代LD50法,更有利于推广、实施。

Objective A new Fluorescence Focus Units（FFU） developed for Rabies Virus titration replace current reference method（LD50） so as to reduce the cost.MethodsThe BSR cell was infected by triple serial dilution of the Rabies Virus CTN strain,then detect the fluorescent focus by FITC labeling anti-Rabies Virus monoclonal antibodies.All the samples were assayed in both FFU and LD50.ResultsData generated indicateed that a significant correlation between the two methods（r=99）.ConclusionThis method is simple and rapid.It will be considered as a useful alternative method to the LD50.