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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 fluorescence quantitative polymerase chain reaction (RT-PCR) for the detection of HIV vaccine against adenovirus and to evaluate the biodistribution of the vaccine in C57BL / 6 mice. Methods: The gag gene fragment was used as the detection object to establish absolute quantitative PCR method. C57BL / 6 mice were intramuscularly injected with 5 × 109vp HIV vaccine intramuscularly, and the tissues were collected on days 3, 15, 30, 60, and 85 after administration. The quantitative real-time quantitative PCR assay was used to detect HIV vaccine. Results: The linear range of detection method was 1 × 102 ~ 1 × 107copies.μL-1. The specificity and repeatability were good. The HIV vaccine is mainly distributed at the site of injection, with a small distribution of lymph nodes, spleen and liver; the amount of vaccine gradually decreases with time. CONCLUSIONS: The intramuscular injection of HIV vaccine resulted in a broad distribution with a time-dependent reduction in the amount of distribution without the risk of germ cell metastasis and the HIV vaccine was safe in mice.