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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 To detect the virulence of rabies virus by replacing 50% Lethal Does (LD50) in mice with Fluorescence Focus Units (FFU) Assay and optimize and improve the existing detection methods to reduce Test costs. Methods Rabbit golden hamster kidney cells were infected by 3-fold serial dilution of rabies virus. The fluorescently-labeled anti-rabies virus nucleoprotein monoclonal antibody was used to detect the fluorescently infected foci. At the same time, the virus was intracranially inoculated with 10 ~ 12g Kunming Mice, by observing the number of mice recorded the incidence of death, counting half the number of mice caused by the death of the virus dose or LD50. Results There was a certain quantitative relationship between FFU and LD50, and there was a positive correlation with a correlation coefficient of 0.99. lgFFU value and LD50 values were statistically significant, good repeatability. Conclusion FFU in vitro titration of rabies virus virulence method can replace the LD50 method, more conducive to the promotion and implementation.