狂犬病病毒不同毒株的生物学特性研究

发布时间：2018-12-17 01:13

【摘要】：目的研究狂犬病不同固定毒株对动物的致病性和对细胞的感染性。方法以小鼠脑内和肌内途径接种比较不同毒株的致病性,研究并建立HEPES(4-羟乙基哌嗪乙磺酸)诱导的空斑形成技术测定病毒空斑形成单位(plaque-forming unit,PFU)和细胞病变感染技术测定病毒感染滴度(CCID50),并用以比较不同毒株的病毒滴度,实验同时以常规的直接免疫荧光法(direct immunofluorescent assay,dFA)做对照。结果不同固定毒株对10~12g小鼠的脑内致病力普遍很高,但肌内毒力普遍较低,脑腔感染致病力比肌内感染致病力高3.0~5.0lg LD50,CVS株相差最大为5.0lg LD50。用两种新的方法 (PFU和CCID50)测定不同毒株的病毒滴度与dFA法测定的结果比较,除个别株外无明显差异,如CVS-11、4aG、PV株用三种方法测定的滴度均在7.1~7.9lg。结论用dFA法测定病毒滴度的结果与小鼠脑内测定的结果有可比性,用以替代小鼠脑内法测定病毒滴度是可行的。两种细胞感染法测定的病毒滴度操作简便,无需贵重仪器和昂贵试剂,可以更广泛地应用于狂犬病病毒和疫苗发展的研究。
[Abstract]:Objective to study the pathogenicity and cell infectivity of different fixed rabies strains to animals. Methods the pathogenicity of different virulent strains was compared by intracerebral and intramuscular inoculation in mice, and the plaque formation technique induced by HEPES (4-hydroxyethylpiperazine ethanesulfonic acid) was established for the determination of viral plaque forming units (plaque-forming unit,). PFU) and cytopathic infection assay (CCID50) were used to compare the viral titers of different virulent strains, and the conventional direct immunofluorescence assay (direct immunofluorescent assay,dFA) was used as control. Results the intracerebral pathogenicity of different fixed strains was very high in 10g / 12g mice, but the intramuscular virulence was generally low. The difference of the pathogenicity of intracerebral infection with 3.0~5.0lg LD50,CVS strain was that of 5.0lg LD50.. Two new methods (PFU and CCID50) were used to determine the virus titers of different strains. There was no significant difference between the two methods except individual strains, for example, the titers of CVS-11,4aG,PV strains measured by the three methods were all 7.1g or 7.9 lg. Conclusion the results of virus titer determination by dFA method are comparable with those of mice brain, and it is feasible to replace the virus titer assay in mice brain. The virus titers determined by two cell infection methods are easy to operate and can be widely used in the research of rabies virus and vaccine development without expensive instruments and expensive reagents.
【作者单位】：中国食品药品检定研究院虫媒病毒疫苗室;
【基金】：国家高技术研究发展计划(863计划)项目(2012AA02A402)
【分类号】：R373.9

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