论文部分内容阅读
目的建立人源轮状病毒的小型猪腹泻模型,为人源轮状病毒感染的免疫保护机制、致病机制及疫苗的研制奠定基础。方法ELISA与RT-PCR相结合分离鉴定野生型人源轮状病毒;用野生型人源轮状病毒G1与G3血清型口服接种不同日龄的小型猪,观察粪便排泄等临床症状,ELISA及免疫荧光分别检测粪便与小肠上皮组织中的轮状病毒抗原,光镜及透射电镜观察小肠上皮组织的病理变化与病毒样颗粒。结果3~5日龄小型猪均出现了典型的临床腹泻症状;粪便中轮状病毒抗原排泄可平均持续5~7 d;小肠组织可见明显的病理变化,同时用免疫荧光检测方法可在小肠组织中检测到特异性轮状病毒抗原;透射电镜进行观察,在小肠组织的超薄切片中可见到大量的轮状病毒颗粒。30~35日龄小型猪可出现明显腹泻临床症状,粪便中轮状病毒抗原排泄可持续4~7 d;56~60日龄小型猪接种野生型人源轮状病毒G1血清型后,均未出现典型的腹泻症状,但可持续排毒2~3 d。结论小型猪可作为人源轮状病毒的理想腹泻动物模型。
Objective To establish a miniature porcine diarrhea model of human rotavirus, which lays the foundation for the development of immunoprotection mechanism, pathogenesis and vaccine of human rotavirus infection. Methods The wild-type human rotavirus was isolated and identified by ELISA and RT-PCR. The mini-pigs of different ages were orally inoculated with the wild-type human rotavirus G1 and G3 serotypes to observe the clinical symptoms, Fluorescence detection of stool and small intestine epithelial tissue rotavirus antigen, light microscopy and transmission electron microscopy of intestinal epithelial tissue pathological changes and virus-like particles. Results The typical clinical symptoms of diarrhea occurred in 3-5 days of miniature pigs. The excretion of rotavirus antigen in feces could last for 5-7 days on average. Significant pathological changes were observed in small intestine. Immunofluorescence assay could be used in small intestine Detected specific rotavirus antigen; transmission electron microscopy was observed in the ultrathin section of intestinal tissue can be seen in a large number of rotavirus particles. The clinical symptoms of diarrhea in mini-pigs aged 30-35 days were obvious. The excretion of rotavirus antigen in the feces was sustainable for 4-7 days. After the 56- to 60-day-old mini-pigs were vaccinated with the wild-type human rotavirus G1 serotype, A typical diarrhea symptoms, but sustainable detoxification 2 ~ 3 d. Conclusion Miniature pigs can be used as an ideal diarrhea animal model of human rotavirus.