原位杂交检测鹅细小病毒VP3基因_省略_P3基因疫苗在小鼠体内的动态分布_李万奎.pdfVIP

888 　 　2010 　30 　 　ChinJ Vet Sci 　Jul.2010　Vol.30 　No .　 VP3 VP3 1,2 1,2 ＊ 1,2 ＊ 1,2 2 1 1,2 (1. 　　李万奎 ,程安春 , 汪铭书 , 朱德康 , 罗启慧 , 陈孝跃 , 范　波 　 , 625014;2., 625014) :将100 μg pcDNA-GPV-VP3 基因疫苗肌肉注射4 周龄BALB/c 鼠, 并以生理盐水和空载体质粒 pcDNA3. 1(+)为对照, 建立并应用寡核苷酸探针原位杂交方法检测pcDNA-GPV-VP3 在免疫 鼠各组织器官(心、肝、脾、 肺、肾、肠、脑、免疫部位肌肉)的动态分布规律。结果表明, 所建立的原位杂交方法具有特异性强、敏感性高、定位 准确等特点,能检测约13 pg 的同源DNA。pcDNA-GPV-VP3 基因疫苗在肌注后1 h 可分布到除脑以外的所有组 织, 肌注后3 h 脑呈阳性, 21 d 除脑外的其他组织仍呈阳性, 105 d 仍能在肌注部位检测到 pcDNA-GPV-VP3;pcD- NA-GPV-VP3在 鼠各组织中持续时间为肌注部位肝脾、心肾肺、肠脑。 :原位杂交;鹅细 病毒VP3 基因疫苗; 鼠;动态分布 :S852.65　　　:A　　　:1005-4545(2010)0 -0888-06 Establishment and application of in situ hybridization for detecting distribution of GPV-VP3 vaccine in mice immunized intramuscularl injection 1,2 1,2 ＊ 1,2 ＊ 1,2 2 　LI Wan-kui , CHENG An-chun ,WANG M ing-shu ,ZHU De-kang , LUO Qi-hui ,CHEN Xi- 1 1,2 ao-yue ,FAN Bo 　(1.Collegeof Veterinary Medicine, Sichuan Agricultural University , Ya ′an, S ichuan 625014, China;2.Key L aboratory of Animal D isease and H uman H ealth of S ichuan Province,Ya ′an,S ichuan 625014, China) Abstract:Three groups of 4-week-old BALB/c mice w ere injected intramuscularly with 100 μg pcDNA-GPV-VP3 gene vaccine, and with normal saline, pcDNA3.1(+)as contro1, in situ hybridization was established and applied to detect the dist ribution of pcDNA-GPV-VP3 in the organs (heart, liver, spleen, lung, kidney, brain, intestine and cutis of immue site)at different time points post-injection.The results showed that the established in situ hybridization was specific, highly sensitive and exactly locative, and can detect 13 pg homology DNA;the pcDNA-GPV-VP3 could be detected in all tissues except brain at 1 h post administration and in brain at 3 h;a