伪狂犬病毒变异株与经典株滴鼻感染小鼠后时空分布规律的研究

发布时间：2018-05-18 03:42

本文选题：伪狂犬病毒(PRV) + 昆明鼠　；参考：《华中农业大学》2017年硕士论文

【摘要】：近年来,伪狂犬病毒(Pseudorabies Virus,PRV)变异株的出现导致原有病毒疫苗的保护力下降,国内猪场伪狂犬病不断爆发,对养猪业造成了巨大的经济损失。本研究以PRV变异株(LY-2015)与经典株(Halv)为研究对象,滴鼻感染成年健康昆明鼠。通过免疫组织化学染色、原位杂交染色、PCR技术结合组织病理学技术,研究病毒在小鼠脑内的分布。同时通过不同时间点小鼠脑内病毒的分布差异分析PRV滴鼻感染小鼠后的上行传导通路,以填补变异株PRV感染小鼠后病毒在脑内分布相关研究的空白,并为临床上PRV的诊断与精确的阻断病毒传播提供基础资料。论文主要研究内容如下:1病毒感染小鼠后的临床症状在2,000TCID50的LY-2015株PRV感染组,小鼠在病毒感染48h后开始出现被毛粗乱、食欲不振;在病毒感染60h后,小鼠表现出精神振奋,抓挠头面部皮肤;病毒感染84h后,多数小鼠因瘙痒抓挠致皮肤破溃,小鼠死亡。用相同感染量的Halv株PRV滴鼻感染小鼠时,未见任何临床症状。在10,000TCID50的Halv株PRV感染组,小鼠在病毒感染72h后开始出现与LY-2015株PRV感染组相似的临床症状;在病毒感染96h后,大量小鼠死亡。结果表明LY-2015株PRV毒力较Halv株PRV强。2病毒感染后的上行传导通路LY-2015株PRV感染小鼠后,病毒首先出现在鼻黏膜下层细胞内,然后在三叉神经相关神经核团、交感神经相关神经核团、面神经相关神经核团内均检测到PRV。表明变异病毒LY-2015株PRV滴鼻感染小鼠后可沿三叉神经、交感神经与面神经通路上行传导,致中枢感染。Halv株PRV滴鼻感染小鼠后,在三叉神经相关核团、交感神经相关核团、副交感神经相关核团与面神经相关核团内检测到PRV。说明经典病毒Halv株PRV滴鼻感染小鼠后可沿三叉神经、交感神经、副交感神经和面神经上行传导。3病毒感染小鼠后在脑内的分布2,000TCID50的LY-2015株PRV感染小鼠后,病毒多分布于脊髓,脑桥和延髓,大脑内仅在丘脑可见少量病毒。在病毒感染84h后,小鼠脑内的病毒分布最为广泛。10,000TCID50的Halv株PRV感染组,病毒不仅分布于脊髓、脑桥和延髓,大脑中也存在多量病毒。病毒感染小鼠96h后在脑内的分布最为广泛。结果表明,经典病毒Halv株PRV感染小鼠后沿轴突上行传导较变异病毒LY-2015株PRV慢;且LY-2015株PRV的毒力较强,小鼠在病毒感染后过早死亡,仅少量病毒传导至中枢,分布于下丘脑外侧部、丘脑下中间核腹侧部等。此外,在2,000TCID50的LY-2015株PRV感染组,小鼠在病毒感染后120h,IHC染色呈PRV阴性,但ISH染色呈PRV阳性,结果表明小鼠脑内存在病毒基因,但病毒未复制。结果表明,感染PRV后,耐过小鼠呈隐性感染,病毒基因整合于小鼠外周神经细胞核内,但不复制产生新的病毒病毒粒子。以上结果表明,LY-2015株PRV毒力强于Halv株PRV;Halv株PRV只有在高病毒滴度下才能造成小鼠的有效感染。滴鼻感染昆明鼠造成有效感染时,变异病毒LY-2015株PRV感染小鼠后沿三叉神经、交感神经与面神经上行传导,但病毒粒子多分布于延髓和脑桥;经典病毒Halv株PRV感染小鼠后可沿三叉神经、交感神经、副交感神经与面神经上行传导,在小鼠脊髓、脑桥、延髓和大脑中均存在多量的病毒分布。在有效感染中耐过小鼠呈潜伏性感染,不表现出任何临床症状,病毒基因整合到小鼠外周神经细胞核中,但不复制。
[Abstract]:In recent years, the occurrence of Pseudorabies Virus (PRV) variant has resulted in the decrease of the protective ability of the original virus vaccine. The domestic swine farm pseudorabies continues to burst out, causing huge economic losses to the pig industry. This study uses PRV variant (LY-2015) and the classic strain (Halv) as the research object, and infect adult healthy Kunming mice by nose drops. Immunohistochemical staining, in situ hybridization, PCR technique combined with histopathological techniques to study the distribution of virus in the brain of mice. At the same time, through the difference of the distribution of virus in the brain of mice at different time points, the uplink pathway of PRV nose drops infected mice was analyzed in order to fill the related research on the distribution of the virus in the brain of the mutant PRV infected mice. The main research contents are as follows: 1 the clinical symptoms of 1 virus infected mice were in the 2000TCID50 LY-2015 strain PRV infection group, and the mice began to be hairy and inexorable after the virus infected 48h; after the virus infected 60H, the mice showed sperm. God exhilarated and scratched the skin of the head and face; after the virus infected 84h, most mice were caused by itching and scratching the skin to break, and the mice died. No clinical symptoms were found when the mice infected with the Halv strain PRV of the same infection rate had no clinical symptoms. In the PRV infection group of Halv strain of 10000TCID50, the mice began to appear similar to the PRV infection group of LY-2015 strain after the virus infected 72h. After the virus infected 96h, a large number of mice died. The results showed that the virulence of LY-2015 strain PRV was more than that of the Halv strain PRV strong.2 virus infection, LY-2015 strain PRV infected mice, the virus first appeared in the submucosa of the nasal mucosa, and then in the trigeminal nerve related nucleus, sympathetic nerve nucleus, and facial nerve related. In the nucleus of the nucleus, PRV. showed that the mutant virus LY-2015 strain PRV was infected with the trigeminal nerve and the sympathetic and facial nerve pathway, causing the central infection of the.Halv strain PRV nose drops in mice, in the trigeminal correlate nucleus, the sympathetic correlate nucleus, the parasympathetic nerve related nucleus and the facial nerve related nucleus. PRV. showed that the classic virus Halv strain PRV infected mice could be infected with the trigeminal nerve, sympathetic, parasympathetic, and facial nerve in the brain, and the LY-2015 strain PRV infected mice in the brain, and the virus was distributed in the spinal cord, the brain bridge and the medulla, and the brain only found a small amount of disease in the thalamus. After the virus infection of 84h, the virus in the brain of the mouse is distributed in the most extensive.10000TCID50 Halv strain PRV infection group. The virus is not only distributed in the spinal cord, the pontine and medulla, but also in the brain. The virus infected mice after 96h is most widely distributed in the brain. The results show that the classical virus Halv strain PRV infected mice along the axon along the axon. The LY-2015 strain was slower than the variant virus strain PRV, and the virulence of LY-2015 strain PRV was stronger. The mice died prematurely after the virus infection, only a small amount of virus was transmitted to the central part of the hypothalamus and the ventral part of the hypothalamus. In addition, in the LY-2015 strain PRV infection group of 2000TCID50, the mice were PRV negative after the virus infection and 120h, but ISH, but ISH, but ISH. The results showed that the PRV positive staining showed that the virus gene existed in the mouse brain, but the virus was not replicated. The results showed that after PRV infection, the mice had the latent infection, the virus gene was integrated in the peripheral nucleus of the peripheral nerve of mice, but no new virus particles were produced. The results showed that the PRV toxicity of LY-2015 strain was stronger than that of Halv strain PRV; Halv strain PRV only. The virus LY-2015 strain LY-2015 PRV infected mice along the trigeminal nerve and the sympathetic and facial nerve conduction, but the virus particles are mostly distributed in the medulla and the pontine; the classic virus Halv strain PRV infected mice can be infected along the trigeminal nerve and sympathetic. The nerve, parasympathetic and facial nerve conduction, in the mouse spinal cord, the pontine, the medulla and the brain all have a large number of virus distribution. In the effective infection, the mice showed latent infection and did not show any clinical symptoms. The virus gene was integrated into the peripheral nerve cell nucleus of the mice, but did not replicate.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S852.65

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