原位杂交检测鹅细小病毒VP3基因方法建立及肌注VP3基因疫苗在小鼠体内的动态分布  

Establishment and application of in situ hybridization for detecting distribution of GPV-VP3 vaccine in mice immunized intramuscularl injection

作　　者：李万奎[1,2] 程安春[1,2] 汪铭书[1,2] 朱德康[1,2] 罗启慧[2] 陈孝跃[1] 范波[1,2] 

机构地区：[1]四川农业大学动物医学院,四川雅安625014 [2]动物疫病与人类健康四川省重点实验室,四川雅安625014

出　　处：《中国兽医学报》2010年第7期888-893,共6页Chinese Journal of Veterinary Science

基　　金：教育部"长江学者和创新团队发展计划"资助项目(PCSIRT0848);现代农业产业技术体系建设专项资金资助项目(nycytx-45-12)

摘　　要：将100μgpcDNA-GPV-VP3基因疫苗肌肉注射4周龄BALB/c小鼠,并以生理盐水和空载体质粒pcDNA3.1(+)为对照,建立并应用寡核苷酸探针原位杂交方法检测pcDNA-GPV-VP3在免疫小鼠各组织器官(心、肝、脾、肺、肾、肠、脑、免疫部位肌肉)的动态分布规律。结果表明,所建立的原位杂交方法具有特异性强、敏感性高、定位准确等特点,能检测约13pg的同源DNA。pcDNA-GPV-VP3基因疫苗在肌注后1h可分布到除脑以外的所有组织,肌注后3h脑呈阳性,21d除脑外的其他组织仍呈阳性,105d仍能在肌注部位检测到pcDNA-GPV-VP3;pcD-NA-GPV-VP3在小鼠各组织中持续时间为肌注部位>肝>脾、心>肾>肺、肠>脑。Three groups of 4-week-old BALB/c mice were injected intramuscularly with 100 μg pcDNA-GPV-VP3 gene vaccine,and with normal saline,pcDNA3.1（＋） as contro1,in situ hybridization was established and applied to detect the distribution of pcDNA-GPV-VP3 in the organs （heart,liver,spleen,lung,kidney,brain,intestine and cutis of immue site） at different time points post-injection.The results showed that the established in situ hybridization was specific,highly sensitive and exactly locative,and can detect 13 pg homology DNA;the pcDNA-GPV-VP3 could be detected in all tissues except brain at 1 h post administration and in brain at 3 h;at 21 d post administration,all tissues were positive except brain,and cutis of injection site were aslo positive after 105 d post administration;the duration time in all tissues was cutis of injectione siteliverheart,spleenkidneylung,intestinebrain.The results demonstrated that pcDNA-GPV-VP3 could distribute in all tissues of mice at l h post administration and existe in injectione site at least 105 d post administration.

关 键 词：原位杂交 鹅细小病毒VP3基因疫苗 小鼠 动态分布 

分 类 号：S852.65[农业科学—基础兽医学]