牛病毒性腹泻病毒E2截短基因重组卡介苗安全性和最佳免疫剂量的研究  

Studies on Safety and Optimal Immune Dose of BVDV-E2 Truncated Gene RBCG

作　　者：杨宇航[1] 李晶[1] 张云[1] 郝俊伟[1] 刘东旭 时坤[1] 李健明[1] 曾范利[1] 杜锐[1,2] 

机构地区：[1]吉林农业大学,吉林长春130118 [2]教育部动物生产及产品质量安全重点实验室,吉林省药用动物二级实验室,吉林长春130118

出　　处：《动物医学进展》2014年第7期34-37,共4页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31372436);国家科技支撑计划项目(2011BAI03B02-1);吉林省科技创新人才培育计划项目(20130521023JH);吉林省科技成果转化促进计划项目(20125067)

摘　　要：将6组BVDV-E2截短基因重组卡介苗用生理盐水稀释,分别进行革兰染色和伊红美蓝平板检测,6组重组卡介苗均无杂菌污染。再对6周龄～8周龄雄性豚鼠进行免疫接种,每只腹腔注射1mL（5×106 cfu/只）,免疫接种豚鼠无任何症状,精神、食欲、排便及发育等均不受影响,且无死亡,说明重组病毒对试验豚鼠是安全的。使用BVDV抗体检测试剂盒对60头健康牛的母源抗体含量进行检测,不含母源抗体占85.00%。取重组卡介苗中的一组,用生理盐水稀释,使重组卡介苗达到105、106、107、108、109 cfu/mL共5组,每组不含母源抗体的牛3头,免疫接种前对每头牛进行尾颈静脉采血,析出血清,用BVDV抗体检测试剂盒测牛血清中抗体水平。采完血后对牛进行皮下免疫接种,4周后,采取血液,检测牛血清中抗体水平。通过免疫接种后的检测结果可知,重组疫苗对牛最佳免疫剂量为108 cfu/mL,为进一步进行重组疫苗回归本动物的免疫效果研究提供了可靠依据。In this experiment,6groups of BVDV-E2truncated gene recombinant BCG which had been diluted with normal saline were tested by Gram stain and eosin methylene blue plate detection respectively,the result showed no bacterial infection.Then the 6-8-week-old male Guinea pigs were immunized intraperitoneally with 1mL（5×106 cfu）recombinant BCG each one.All of the Guinea pigs had no any systemic symptoms and their spirit,appetite,feces and growth etc.were not affected,and no deaths,which proved that the recombinant BCGs were safe to the Guinea pigs.The levels of maternal antibodies of 60 healthy cows were detected with the BVDV antibody detection kit and the proportion that had no maternal antibodies was 85.00%.Then one group of recombinant BCG was diluted with saline to make the amount of recombinant BCG to 105,106,107,108,109 cfu/mL respectively.Subsequently,three 24-month-year-old cows which had no maternal antibodies were immunized with each dose.Before immunization,every cow＇s blood were collected from tail vein,for testing the level of antibody in the serum with the BVDV antibody detection kit.The subcutaneous immunization was made after the blood was collected.Four weeks later,the blood was collected again for seral antibody detection.The result showed that the optimal immune dose of recombinant vaccine was 108 cfu/mL,which provided a reliable basis to study the immune effect of the recombinant vaccine when it was applied to the original animals.

关 键 词：牛病毒性腹泻病毒 E2截短基因 重组卡介苗 最佳免疫剂量 安全性 

分 类 号：S852.659.6[农业科学—基础兽医学]