摘要
目的开展猪轮状病毒OSU株的细胞培养特性及致病性研究,确定轮状病毒培养的关键技术与致病规律。方法以MA104细胞培养病毒,对预处理病毒的胰酶浓度与时间、维持液中最佳胰酶浓度等关键条件进行优化,透射电镜观察病毒粒子形态,测定病毒TCID50,口服感染3d龄仔猪进行致病性试验。结果病毒经20μg/mL胰酶预处理1h,37℃吸附细胞2h,维持液最佳胰酶浓度为4μg/mL,病毒典型细胞病变为病变细胞葡萄串状堆积、胞浆相连、拉网等病变。透射电镜下病毒粒子呈圆形车轮状,直径约80nm。病毒感染3日龄仔猪10h后出现典型的黄色水样腹泻,感染42h后死亡,剖检可见胃内有凝乳块、肠壁变薄充满液体,盲肠、结肠充气。主要病变为:肠上皮细胞变性、坏死、脱落;固有膜高度扩张、充血和伴有轻微出血;粘膜上皮脱落,粘膜下层水肿、炎性细胞浸润等。结论研究阐明了轮状病毒OSU株的培养特性与致病特征。
Porcine rotavirus was cultured successfully in MA104 cells. The virus was treated for lh by trypsin on final concentration of 20μ/mL, and then inoculated in maintain medium with 4μ/mL trypsin. The cytopathic effects such as kytoplasm connecting, botryoidalis accretion and netballoon were clearly observed. Double capsid virus was detected by electronmi-croscope. Healthy 3-dayold colostrumsdeprived piglets were inoculated orally with 1 ×10（6.8） TCID_50 of porcine rotavirus. Piglets developed watery diarrhea 10h post infection, and died in 42h post infection. During necropsy, milk curd in stomach, fluid in thinwalled small intestines, and ectocolon were observed. During histopathological, examination, sight to moderate viilous stunting were observed. Bulging and sloughing of villous epithelial cells were prominent. Neutrophils were clustered in the lamina propria at villous tips subjacent to disrupted epithelium. In severity segment of villous destruction, necrosis and sloughing of exposed lamina propria resulted in complete loss of villous strcture.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2012年第2期120-123,147,共5页
Chinese Journal of Zoonoses
基金
国家高技术研究发展计划(863计划)(2009AA10Z402)
国家农业部公益性行业(农业)科研(201203056)
教育部<长江学者和创新团队发展计划>创新团队项目(IRTO848)联合资助
关键词
猪轮状病毒
培养特性
人工感染
致病性
porcine rotavirus
cell culture,
experimental infection
pathogenicity
