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当前位置: 首页 > 文章 > 基因枪轰击不同剂量小鹅瘟病毒VP3基因疫苗在雏鹅体内的动态分布 畜牧兽医学报 2007,38 (11) 1204-1210
Position: Home > Articles > Dynamic Distribution of GPV-VP3 Gene Vaccine in Geese Vaccinated with Gene-gun Bombardment Chinese Journal of Animal and Veterinary Sciences 2007,38 (11) 1204-1210

基因枪轰击不同剂量小鹅瘟病毒VP3基因疫苗在雏鹅体内的动态分布

作  者：
黎敏;程安春;汪铭书;韩新锋;刘晓东;卢菲;车茜;陈孝跃
单  位：
四川农业大学动物医学院禽病防治中心
关键词：
FQ-PCR;pcDNA-GPV-VP3;基因枪;雏鹅;动态分布
摘  要：
本文开展了检测小鹅瘟病毒(GPV)VP3基因的实时荧光定量PCR(FQ-PCR)方法的建立和基因枪轰击不同剂量(1、3和6μg)GPV-VP3基因疫苗(pcDNA-GPV-VP3)在30日龄四川白鹅体内(心、肝、脾、肺、肾、法氏囊、胸腺、哈氏腺、十二指肠、空肠、回肠、直肠、盲肠、胰腺、血液、脑及注射部位皮肤)分布规律的研究。结果表明:①建立的FQ-PCR特异性强、灵敏度高、重复性好,核酸模板数与FQ-PCR测定的Ct值相关系数达到0.999,具有很好的直线相关性;②pcDNA-GPV-VP3各剂量免疫雏鹅1 h即可在各组织中检测到,其中注射部位含量最高,肝、肾、淋巴器官(脾、法氏囊、胸腺、哈氏腺)含量较高;③到免疫后217 d时,1μg组免疫雏鹅各个组织器官内仍检测到pcDNA-GPV-VP3的存在,但多数组织器官中的含量比1 h时约少了4个数量级,其中免疫部位减少了7个数量级;④血液中pcDNA-GPV-VP3的含量较少,且免疫后1 h~217 d各时间点的差异不显著(P≥0.05);⑤不同剂量pcDNA-GPV-VP3免疫雏鹅各组织中的含量呈现的总体规律为6μg组>3μg组>1μg组,但差异不显著(P≥0.05)。因此,FQ-PCR是定量检测pcDNA-GPV-VP3在免疫雏鹅体内含量的可靠方法,pcDNA-GPV-VP3免疫雏鹅后1 h时可分布至雏鹅体内各组织器官中并持续存在217 d以上。
译  名：
Dynamic Distribution of GPV-VP3 Gene Vaccine in Geese Vaccinated with Gene-gun Bombardment
作  者：
LI Min1,CHENG An-chun1,2 ,WANG Ming-shu1,2 ,HAN Xin-feng1,LIU Xiao-dong1,LU Fei1,CHE Qian1,CHEN Xiao-yue1,2 (1.Avain Disease Research Center,College of Veterinary Medicine,Sichuan Agricultural University,Yaan 625014,China;2.Key Laboratory of Animal Disease and Human Health of Sichuan Province,Yaan 625014,China)
关键词：
FQ-PCR;pcDNA-GPV-VP3;gene-gun;geese;dynamic distribution
摘  要：
This paper concentrates on developing the Real-time PCR method for detecting GPV-VP3 gene and studying the dynamic distribution of GPV-VP3 gene vaccine(pcDNA-GPV-VP3)in geese(duodenum,liver,spleen,kidney,bursa of Fabricius,thymus,ileum,cecum,pancreas,heart,blood,jejunum,lung,gland of Harder,skin of injected spot,rectum and brain).Ninety 30-day-old geese were divided into 5 groups.Animals in pcDNA-GPV-VP3 inoculation groups were injected via gene-gun with different doses(1,3,6 μg),respectively.Results showed that: ①This assay was specific,highly sensitive and rapid for detecting pcDNA-GPV-VP3.The standard curve showed a good linear relationship between Ct and template concentrations,and the correlation coefficient was 0.999.②The pcDNA-GPV-VP3 could be detected in all tissues and peripheral blood 1 h post immunization.The copy numbers were most at the skin of injected spots,then liver,kidney,lymphoid organ(spleen,bursa of Fabricius,thymus,gland of Harder).③After 217 d post-administration,pcDNA-GPV-VP3 could still be detected in tissues of 1 μg dose group.But the copy numbers decreased about 104 copies in the most tissues than that at 1 h post administration,the obvious decrease of pcDNA-GPV-VP3 was found at the injection sites(it dropped about 107 copies).④The copy numbers in the serum were always little and changed not obvious with the time(P≥0.05).⑤Different dosage groups of pcDNA-GPV-VP3 distributed in the geese tissues did not significantly differ from each other(P≥0.05),the 6 μg group had the highest copy numbers,then the 3 μg group,and the 1 μg group had the lowest.The results demonstrated that FQ-PCR was a reliable method for detecting dynamic distribution of pcDNA-GPV-VP3,and pcDNA-GPV-VP3 could distribute in all tissues of geese 1 h post immunization,and exist in those at least 217 days post immunization.

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