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。
Objective To study the pathogenicity and infectivity of different rabies virus strains in mice and cell cultures. Methods Mice （10-12 g） were inoculated intracerebrally or intramuscularly with six established virus strains, and observed the mortality within 14 days. HEPES enhancing plaque assay was used to measure the virus plaque forming unit （PFU）, and the CPE stained assay to measure CCID50 in BHK-21 and Veto cells. Two new methods were compared to detect the virus infectivity with routine dFA assay. Results All 6 strains were highly pathogenic to the mice by intracerebral inoculation but weaker pathogenic by intramuscular inoculation. The pathogenicity of intracerehral inoculation was 3.0-5.0 lg LD50 which is higher than that of intramuscular inoculation. The result of dFA assay was comparable to that routinely used for virus titration in vaccine production. The infectivity tested using the new methods, PFU and CPE, was comparable to that of the routinely used dFA method. Conclusions The dFA method can be used to substitute the mouse intracerebral inoculation to evaluate the titration of virus in the vaccine quality control. The two new methods were technically easy without expensive instruments and reagents and thus can be applied in the study of rabies virus and rabies virus vaccine.
出处
《中国病毒病杂志》
CAS
2014年第3期179-186,共8页
Chinese Journal of Viral Diseases
基金
国家高技术研究发展计划(863计划)项目(2012AA02A402)
