论文部分内容阅读
目的 研究呼吸道合胞病毒 (RSV)感染模型的建立。方法 实验组用在HEP 2细胞上培养出的不同滴度 (7 5× 10 5,2 75× 10 6,2 5× 10 7PFU/ml)的 9815 9株RSV予Balb/c小鼠滴鼻 ,每个滴度 3只小鼠 ,共 9只。对照组 3只小鼠用HEP 2细胞悬液滴鼻 ,5d后 ,无菌状态下取肺 ,右肺称重 ,按1∶10加入eagleMEM ,匀浆 ,4℃ ,2 0 0 0r/min离心 2 0min ,上清液加入HEP 2细胞作病毒分离 ,空斑分析。左肺用于病理检查 ,电镜检查。结果 (1)RSV感染使梭形的HEP 2细胞融合成地图状、圆形、椭圆形、树枝状 ;(2 )肺组织匀浆接种于HEP 2细胞上 ,于接种的 10～ 12d分离出病毒 ;(3)不同滴度的RSV在肺组织中的复制滴度不同 ,滴度越高 ,肺复制滴度越高 ;(4 )较高滴度的RSV感染小鼠 ,肺组织病毒复制于感染后的 4～ 6d达高峰 ,肺病理变化于 5～ 8d最明显 ,可见炎症细胞浸润 ,主要是淋巴细胞 ;低滴度RSV感染小鼠后病理变化不明显 ;(5 )电镜检查 :高倍镜下可见毛刺样外观的病毒 ,病变呈横切或纵切。结论 采用 9815 9株呼吸道合胞病毒滴鼻可成功地在Balb/c小鼠身上复制出RSV肺部感染的模型
Objective To study the establishment of respiratory syncytial virus (RSV) infection model. Methods The experimental group was administered intranasally with 98159 RSV to Balb / c mice with different titer (7 5 × 10 5, 2 75 × 10 6, 25 × 10 7 PFU / ml) cultured on HEP 2 cells, Three mice per titre, 9 in total. Three mice in the control group were intranasally inoculated with HEP 2 cell suspension. After 5 days, the lungs were aseptically removed and the right lungs were weighed. The eagle cells were homogenized at 1:10 and centrifuged at 4 ° C at 200r / min 20min, the supernatant was added HEP 2 cells for virus isolation, plaque analysis. Left lung for pathological examination, electron microscopy. Results (1) RSV infection fused fusiform HEP 2 cells into map-like, round, oval and dendritic shape. (2) The lung tissue homogenate was seeded on HEP 2 cells and virus was isolated 10-12 days after inoculation ; (3) different titers of RSV replication in the lung tissue titer, the higher the titer, lung replication titers higher; (4) higher titers of RSV infection in mice, lung virus replication in the infection After 4 ~ 6d reached the peak, the pathological changes of lungs were most obvious on the 5th ~ 8th day, and inflammatory cell infiltration was observed, mainly lymphocytes; pathological changes were not obvious in low-titer RSV infected mice; (5) Electron microscopy: Burr-like appearance of the virus can be seen lesions were transverse or longitudinal incision. Conclusion 98159 respiratory syncytial virus nasal drops can successfully replicate the model of RSV pulmonary infection in Balb / c mice