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, Vol. 38 ›› Issue (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

黎敏；程安春；汪铭书；韩新锋；刘晓东；卢菲；车茜；陈孝跃

1.四川农业大学动物医学院禽病防治中心，雅安 625014；2.动物疫病与人类健康四川省重点实验室，雅安 625014

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-11-25 发布日期:2007-11-25

Dynamic Distribution of GPV-VP3 Gene Vaccine in Geese Vaccinated with Genegun Bombardment

LI Min；CHENG An-chun；WANG Ming-shu；HAN Xin-feng；LIU Xiao-dong；LU Fei；CHE Qian；CHEN Xiao-yue

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

Received:1900-01-01 Revised:1900-01-01 Online:2007-11-25 Published:2007-11-25

摘要/Abstract

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6 μg）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，1 μg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6 μg组＞3 μg组＞1 μg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。

关键词: FQ-PCR, pcDNA-GPV-VP3, 基因枪, 雏鹅, 动态分布

Abstract: 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.

Key words: FQ-PCR, pcDNA-GPV-VP3, gene-gun, geese, dynamic distribution

黎敏;程安春;汪铭书;韩新锋;刘晓东;卢菲;车茜;陈孝跃. 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[J]. 畜牧兽医学报, 2007, 38(11): 1204-1210.

LI Min;CHENG An-chun;WANG Ming-shu;HAN Xin-feng;LIU Xiao-dong;LU Fei;CHE Qian;CHEN Xiao-yue. Dynamic Distribution of GPV-VP3 Gene Vaccine in Geese Vaccinated with Genegun Bombardment [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2007, 38(11): 1204-1210.

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