鸭特异性CpG ODN体外抑制鸭坦布苏病毒复制的研究  

Inhibitory Effect of Duck Specific CpG ODN on Duck Tembusu Virus Replication in Vitro

作　　者：李海燕[1,2] 于森[1] 刘伟杰[1] 林冬梅[2] 黄艳艳[1] 杨少华[1] 张琳[1] 吴家强[1] 

机构地区：[1]山东省农业科学院畜牧兽医研究所山东省畜禽疫病防治与繁育重点实验室,山东济南250100 [2]河北工程大学生命科学与食品工程学院,河北邯郸056021

出　　处：《中国家禽》2017年第16期17-21,共5页China Poultry

基　　金："十三五"国家重点研发计划课题(2016YFD0500106);"十二五"农村领域国家科技计划项目(2015BAD12B03);山东省农业科学院农业科技创新工程(CXGC2016B14);泰山学者特聘专家工程经费(ts201511069)

摘　　要：为明确鸭特异性寡聚脱氧核苷酸(Cp G ODN)体外对鸭坦布苏病毒(DTMUV)复制的抑制作用,研究在Cp G ODN最佳活性分子筛选的基础上,与DTMUV感染后的细胞进行共孵育,分别采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐法(MTT法)和实时荧光定量PCR(q RT-PCR)检测不同浓度Cp G ODN-D2对DTMUV复制的抑制作用。结果显示:对非洲绿猴肾(Vero)细胞来说,Cp G ODN-D2浓度在300μg/m L以内均为安全浓度;MTT法和q RT-PCR检测结果均表明,各浓度Cp G ODN-D2对DTMUV的复制均有一定的抑制作用,并随浓度升高呈现先上升后下降的趋势,其中30μg/m L的浓度效果最佳,达到84.74%。结果证实了Cp G ODN-D2作为新型免疫调节剂可有效抑制DTMUV的复制。In order to evaluate the inhibitory effect of duck specific CpG oligonucleotide （CpG ODN）on duck Tembusu virus （DTMUV）replication in vitro, the optimal CpG ODN was screened firstly and incubated with cells infected with DTMUV. After 72 hours, the inhibition rate of CpG ODN-D2 proliferation at different concentrations were analyzed by 3-（4, 5- Dimethylthiazol-2- yl）-2, 5-diphenyltetrazolium bromide （MTT）method and quantitative real-time PCR （qRT-PCR）. The results showed that the proliferation rate of Vero cell was still higher than negative control even treated with CpG ODN-D2 at the concentration of 300μg/mL, which indicated the maximum safe concentration was over 300 μg/mL. According to the results of MTT and qRT-PCR, inhibition rate of CpG ODN-D2 upregulated firstly and then downregulated with the dose increasing, but the effect exhibited significantly at all concentrations. The best inhibitory effect on virus proliferation in vitro was 84.74% when the concentration of CpG ODN-D2 was 30 μg/mL. This work demonstrated the in vitro anti-virus effect of CpG ODN-D2 on DTMUV.

关 键 词：鸭 CPGODN DTMUV 抑制 

分 类 号：S855.3[农业科学—临床兽医学]