柯萨奇病毒A16型感染性模型的建立及其相关免疫学指标和应用的评价

发布时间：2018-03-29 06:31

本文选题：柯萨奇病毒A型　切入点：长爪沙鼠　出处：《中国比较医学杂志》2017年01期

【摘要】：目的建立简便和可靠的对柯萨奇病毒A16型(coxsackievirus A16,CVA16)敏感的小型啮齿类实验动物模型。方法选取不同日龄长爪沙鼠,腹腔接种CVA16后连续观察14 d,筛选出对病毒敏感的最适接种日龄段;然后比较接种剂量与效应关系,测定其50%致死剂量(LD50);并测定感染3 d后其血液和主要组织器官中CVA16病毒滴度;最后用CVA16灭活疫苗在1日龄和11日龄免疫2针沙鼠后,在14日龄时用LD50剂量病毒攻毒,观察并记录沙鼠体重、症状及死亡率,2周后眼球采血,应用微量中和试验和ELISA分别检测中和抗体效价和总抗体水平。结果长爪沙鼠感染CVA16后出现活动减少、后肢无力、麻痹瘫痪、死亡等临床症状,不同日龄沙鼠对CVA16感染的易感能力和感染后发病程度不同,7日龄和14日龄沙鼠易感,28日龄沙鼠不易感染,最敏感和最合适接种日龄为14日龄,其LD50为1×104.5 CCID50,感染3 d后其血液和主要组织器官中均可检测到高滴度的CVA16病毒,免疫两针灭活疫苗的14日龄沙鼠用LD50剂量攻毒,存活率87.5%,疫苗免疫诱导沙鼠产生的CVA16中和抗体几何平均值(GMT)为28.14,抗体阳性率为87.5%。结论长爪沙鼠对CVA16敏感并且病毒能在其体内进行有效的复制和繁殖,可作为CVA16致病机制研究、疫苗研发及药物评价的可靠小动物模型。
[Abstract]:Objective to establish a simple and reliable small rodent model sensitive to Coxsackievirus A16 (Coxsackievirus A16). Then, the relationship between inoculation dose and effect was compared, the 50% lethal dose of LD50 was measured, the titer of CVA16 virus in blood and main tissues and organs were measured 3 days after infection, and then, two injections of gerbils were immunized with CVA16 inactivated vaccine at the age of 1 and 11 days. The body weight, symptoms and mortality of gerbils were observed and recorded at the age of 14 days. Blood was collected from the eyeballs of gerbils after 2 weeks. The neutralization test and ELISA were used to detect neutralization antibody titer and total antibody level respectively. Results after infection with CVA16, the activity decreased, hind limb weakness, paralysis, death and other clinical symptoms occurred in gerbils. The susceptibility to CVA16 infection in gerbils of different ages and the degree of morbidity after infection were different. The susceptible gerbils of 7 and 14 days of age were not susceptible to infection, and the most sensitive and suitable inoculation days were 14 days old. Its LD50 was 1 脳 104.5 CCID50. After 3 days of infection, high titer of CVA16 virus could be detected in its blood and main organs. 14-day-old gerbils immunized with two doses of inactivated vaccine were attacked with LD50 dose. The survival rate was 87.5%. The geometric mean value of CVA16 neutralizing antibody produced in gerbil induced by vaccine immunization was 28.14, and the positive rate of antibody was 87.5%. Conclusion the gerbils with long claws are sensitive to CVA16 and the virus can replicate and reproduce effectively in their bodies, which can be used to study the pathogenesis of CVA16. Reliable small animal models for vaccine development and drug evaluation.
【作者单位】：浙江省医学科学院病毒病研究所;
【基金】：浙江省科技计划项目(2014C37014;2016C37102)
【分类号】：R-332;R512.5

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