摘要
将本实验室构建的鸭α-干扰素基因疫苗(pcDNA-SDIFN-α)分别按每只50、100、200μg3个剂量肌肉注射免疫樱桃谷鸭,以PBS、空载体质粒pcDNA3.1(+)和鸭瘟弱毒疫苗为对照,免疫15d后攻击感染鸭瘟强毒,于攻毒后2h、6h、12h、24h、3d、6d、9d、14d、22d、26d、33d和40d采全血,同时取死亡鸭的各组织器官,采用实时荧光定量PCR对鸭瘟病毒在鸭外周血中的动态变化和在各组织器官中的分布及含量进行检测。结果表明:①3个剂量pcDNA-SDIFN-α和鸭瘟弱毒疫苗都对鸭产生良好保护作用,免疫鸭未发生死亡;而PBS和空载体对照组3只鸭中有1只鸭死亡,死亡鸭心、肝、脾、肾、胰和各段肠管中均检测到鸭瘟病毒DNA且其含量大于pcDNA-SDIFN-α免疫鸭外周血病毒DNA含量;②3个剂量pcDNA-SDIFN-α免疫鸭外周血鸭瘟病毒DNA含量比PBS和空载体对照组低,差异显著(P<0.05),特别是在2h差异极显著(P<0.01);攻毒后24h内,3个剂量pcDNA-SDIFN-α免疫鸭外周血鸭瘟病毒DNA含量均低于鸭瘟弱毒疫苗免疫鸭,差异显著(P<0.05);3个不同剂量pcDNA-SDIFN-α免疫组之间外周血鸭瘟病毒DNA含量差异不显著(P>0.05),攻毒初期,200μg免疫鸭外周血鸭瘟病毒DNA含量最低,100μg次之,50μg最高。研究表明,pcDNA-SDIFN-α肌肉注射免疫鸭后能产生一定的抗鸭瘟强毒感染的作用,并在攻毒初期表现出一定的量效关系。
Yingtaogu ducks were immunized once with 50, 100, 200 μg pcDNA - SDIFN - α DNA vaccine respectively by the route of intramuscular injection with PBS, pcDNA3.1( + ). We attenuated duck plague virus vaccine as control, challenged with duck plague virus (DPV) after immunization 15 days later and collected the peripheral blood samples at 2 h, 6 h, 12 h, 24 h, 3 d, 6 d, 9 d, 14 d, 22 d, 26 d, 33 d and 40 d, as well as tissues or organs of dead ducks. The real-time fluorescence quantitative PCR was used to detect the number of DPV - DNA in blood and tissues or organs. Results: ①Ducks of pcDNA-SDIFN-α groups and attenuated DPV vaccine groups were not dead, but one duck of PBS and pcDNA 3.1 ( + ) groups was dead. The DPV - DNA can be detected from heart, liver, spleen, kidney, pancreas and all intestines, and the quantity of DPV- DNA was much more than the peripheral blood of pcDNA - SDIFN - α groups; OThe quantity of DPV - DNA in peripheral blood was significantly (P 〈 0.05 ) or highly significant ( P 〈 0.01 ) different among pcDNA - SDIFN - α groups and PBS group, pcDNA3.1 ( + ) group. The quantity of DPV - DNA in peripheral blood was significantly (P 〈 0.05) different between attenuated DPV vaccine group. The quantity of DPV - DNA in peripheral blood was not different (P〉0.05) between 50, 100μg group and 200μg group. The quantity of DPV- DNA of 200μg group was the least, 100μg group was more, 50μg group was the most. Conclusion, peDNA - SDIFN - α immunized by intramuscular injection can counteract the infection of DPV and show some dose-dependent relationship.
出处
《四川农业大学学报》
CSCD
2007年第4期475-479,共5页
Journal of Sichuan Agricultural University
基金
国家科技攻关重大项目(2004BA901A03)
教育部"新世纪优秀人才支持计划"项目(NCET-04-0906)
四川省教育厅重点项目(2003A024)
四川省重点建设学科项目(SZD0418)
关键词
α-干扰素基因疫苗
荧光定量PCR
鸭瘟病毒
interferon-α gene vaccine
fluorescence quantitative PCR
duck plague virus