鸭α-干扰素基因疫苗在鸭体内动态分布规律的定量检测  

Examination of dynamic distributions of DuIFN-α gene vaccine in ducks by real-time fluorescent quantitative PCR

作　　者：段坤[1] 程安春[1] 汪铭书[1] 周雪[1] 程志萍[1] 刘闯[1] 陈孝跃[1] 陈斌[1] 

机构地区：[1]四川农业大学动物科技学院禽病防治中心,四川雅安625014

出　　处：《中国兽医科学》2007年第5期390-395,共6页Chinese Veterinary Science

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);四川省重大基础研究项目(05JY029-109);四川省重点项目(2003A024);四川省重点建设学科项目(SZD0418)

摘　　要：将50、100和200μgpcDNA—SDIFN—α分别肌肉注射28日龄樱桃谷肉鸭，设PBS和空载体质粒pcDNA3．1（＋）对照，建立并应用TaqMan探针实时荧光定量PCR检测了不同时间pcDNA—SDIFN—α在血液以及各组织器官中的含量。结果，建立的PCR具有特异性强、灵敏度高、快速等特点，各剂量pcDNA—SDIFN—α给鸭免疫后的第1h即可在血液及各组织中检测到，注射部位显著高于除心脏外的其他组织；除血液外其他器官中pcDNA—SDIFN—α的含量在1～24h达到最高，随后逐渐减少；105d时各组织仍能检测到（约1×10^2拷贝／g）。不同剂量pcDNA—SDIFN-α在免疫鸭各组织中的含量的总体规律为200μg〉100μg〉50μg。表明，pcDNA—SDIFN—α肌肉注射能迅速分布到鸭体各组织器官中并持续存在105d以上。Five groups of 28-day-old ducks were injected intramuscularly with 50,100 and 200 μg of pcDNA-SDIFN-α, respectively, with PBS and pcDNA3.1 （＋） as controls. A real-time fluorescent quantitative PCR was established and applied to detect the copy numbers of pcDNA-SDIFN-α in peripheral blood and organs at different time points post-injection（PI）. The established PCR was specific, highly sensitive, and rapid for detecting pcDNA-SDIFN-α. The DNA vaccine pcDNA-SDIFN-α could be detected in all tissues and peripheral blood at hour 1 PI. The copy numbers of pcDNA-SDIFN-α at the injection site was significantly more than those at other samples except the heart. The copy numbers of pcDNA-SDIFN-α at each tissue reached peak at hour 1 to 24 PI, then it began to decrease. However,on day 105 PI,it could still be detected in all the tissues, but the copy numbers were only 1 ×10^2 copies per gram. It was concluded that the DNA vaccine pcDNA-SDIFN-α could be quickly distributed in every tissue after being injected intramuscularly,and it could be in existence for more than 105 d.

关 键 词：实时荧光定量PCR 鸭a-干扰素基因疫苗 鸭 

分 类 号：S852.42[农业科学—基础兽医学]