摘要
目的:建立腺病毒载体HIV疫苗检测的实时荧光定量PCR方法,并评价疫苗在C57BL/6小鼠体内的生物分布。方法:以gag基因片段为检测对象,建立绝对定量PCR方法。C57BL/6小鼠单次肌内注射5×109vp HIV疫苗,给药后d 3,15,30,60,85收集组织脏器,用建立的荧光定量PCR方法定量检测HIV疫苗。结果:检测方法线性范围为1×102～1×107copies.μL-1,特异性强,重复性较好。HIV疫苗主要分布在注射位点,淋巴结、脾脏和肝脏也有少量分布;疫苗的量随时间延长逐渐减少。结论:肌内注射HIV疫苗后导致广泛的分布,分布的量呈时间依赖性减少,没有生殖细胞转移的危险,HIV疫苗在小鼠体内是安全的。
Objective ： To establish a real-time quantitative PCR method for the detection of adenovirus vector HIV vaccine and to evaluate the biodistribution of the vaccine in C57BL/6 mice. Methods：A real-time quantitative PCR method was established according to the gag gene sequences. C57BL/6 mice were inject im with 5 ×10^9 vp HIV vaccine, and collect tissues and organs at 3, 15, 30, 60 d and 85 d after injection. Copy numbers of HIV vaccine were quantified by the real-time quantitative PCR. Results：The linearity range of the method is 1 ×10^2 - 1 ×10^7 copiesμL-1 The specificity and reproducibility were good. HIV vaccines were mainly distributed in injection site, while a small quantity of vaccines was in lymph node, spleen and liver;Quantity of HIV vaccine diminished gradually as time goes on. Conclusion：The HIV vaccine has wide distribution which decreases with time. No potential of germ-line transmission has been shown. The vaccine is safe in mice.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2012年第10期1085-1087,1098,共4页
Chinese Journal of New Drugs
基金
国家"重大新药创制"科技重大专项(2008ZX09305-002)
