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作 者:刘发志[1] 陈毓川[2] 赵青松[1] 刘方玉[1]
机构地区:[1]三峡大学职业技术学院,宜昌443003 [2]华中农业大学,武汉430070
出 处:《中国预防兽医学报》2001年第3期206-208,共3页Chinese Journal of Preventive Veterinary Medicine
摘 要:用犊牛睾丸细胞培养传染性牛鼻气管炎病毒 (IBRV) ,反复冻融 ,差速离心提取病毒 ,用TritonX_10 0溶解 ,超声波处理 ,制成IBRVTritonX_10 0亚单位抗原 ,经SDS_PAGE电泳 ,其分子量在 176kD~ 5 3kD之间 ,其中有 6条清晰的蛋白带。该抗原与一定比例的白油_司盘佐剂乳化 ,接种育成牛 ,经间接ELISA检测 ,可使接种的育成牛产生高效价的血清抗体 (OD4 92nm =1.5 7)。2种不同剂量 (80mg/头 ,40mg/头 )的IBRV亚单位疫苗接种育成牛 ,体内抗体效价相差不显著 ,抗体可在体内持续 12周 ,用IBRVTritonX_10 0亚单位疫苗 2次接种育成牛 ,其体内抗体效价显著高于用灭活疫苗 2次接种育成牛体内的效价。试验结果表明 :提取的IBRV多肽制作的亚单位疫苗具有良好的免疫原性 ,且抗体持续时间较长。Infectious Bovine Rhinotracheitis Virus (IBRV)infected calf testicular cells for an appropriate time was cultured.The infected cells were used for the preparation of subunit vaccine by free_thawing,treated with nonionic detergent TritonX_100 and supersonic wave.According to the SDS_PAGE electrophoresis there showed 6 clear bands,The vaccine molecular weight would be between 176Kd_53Kd,After emulsifying completely with white oil and span,it was injected to the calves,The vaccine had induced potent immunity.ELISA was performed for antibody titers and showed high titers of it (OD 492nm =1.57),Two different dosages (80mg/head/time or 40mg/head/time)of subunit vaccine could induce almost the same titers.The antibody.lasted about 12 weeks,while the inactivated vaccine of IBRV only lasted about 6 weeks.Calves vaccinated with the subunit vaccine had obviously higher antibody titers than that of inactivated vaccine after calves were inoculated against IBRV subunit vaccine or inactivated vaccine in two times.Results showed that IBRV subunit vaccine have better antigenicity and can last for a longer period of immunity.
关 键 词:传染性牛鼻气管炎病毒 亚单位疫苗 间接ELISA 免疫原性
分 类 号:S858.236.3[农业科学—临床兽医学] S852.653[农业科学—兽医学]
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