关键词: 荧光定量PCR, 小鹅瘟病毒vp3基因疫苗, 基因枪, 小鼠, 动态分布

Abstract: In this paper we established the method of Fluorescent quantitative PCR (FQ-PCR) for detecting VP3 gene of Gosling plague virus (GPV) and studied the dynamic distribution of GPV-VP3 gene vaccine (pcDNA-GPV-VP3) in BALB/c mice tissues (cardiac muscle, liver, spleen, lung, kidney, brain, intestine and cutis of immune site) vaccinated via gene gun bombing with different dose (6 μg per mouse, 3 μg per mouse and 1 μg per mouse). The results show ed that:①The method of FQ-PCR is specific, sensitive and good repeated. There was a good linear correlation between the copy numbers of the nucleic acid and Ct value detected by FQ-PCR (correlation coefficients=0.999);②pcDNA-GPV-VP3 was detected in all tissues 1 h post-inoculation. The copy numbers of pcDNA-GPV-VP3 was largest in cutis of immune site, and very high in cardiac muscle and lung, and the least in brain;③The copy numbers of pcDNA-GPV-VP3 in tissues began to decrease 3 h post-inoculation. pcDNA-GPV-VP3 was still detected in all tissues 31 wk post-inoculation, but the copy numbers decreased about 10² in most tissues and about 10³ in cutis of immune site than 1 h tissues;④ The ranking of the copy numbers of pcDNA-GPV-VP3 in the same tissues was 6 μg per mouse > 3 μg per mouse >1 μg per mouse, but the difference among them was not significant (P>0.05). The research demonstrated that FQ-PCR was a reliable experiment tool for detecting the copy numbers of pcDNA-GPV-VP3 in mice, and pcDNA-GPV-VP3 could distribute to all over mice bodies 1 h and existed more than 31 wk post-inoculation.

Key words: fluorescent quantitative PCR, gosling plague virus VP3 gene vaccine, gene gun, mice, dynamic distribution