本文選題：偽狂犬病毒(PRV) + 昆明鼠��； 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：近年來,偽狂犬病毒(Pseudorabies Virus,PRV)變異株的出現(xiàn)導(dǎo)致原有病毒疫苗的保護(hù)力下降,國(guó)內(nèi)豬場(chǎng)偽狂犬病不斷爆發(fā),對(duì)養(yǎng)豬業(yè)造成了巨大的經(jīng)濟(jì)損失。本研究以PRV變異株(LY-2015)與經(jīng)典株(Halv)為研究對(duì)象,滴鼻感染成年健康昆明鼠。通過免疫組織化學(xué)染色、原位雜交染色、PCR技術(shù)結(jié)合組織病理學(xué)技術(shù),研究病毒在小鼠腦內(nèi)的分布。同時(shí)通過不同時(shí)間點(diǎn)小鼠腦內(nèi)病毒的分布差異分析PRV滴鼻感染小鼠后的上行傳導(dǎo)通路,以填補(bǔ)變異株P(guān)RV感染小鼠后病毒在腦內(nèi)分布相關(guān)研究的空白,并為臨床上PRV的診斷與精確的阻斷病毒傳播提供基礎(chǔ)資料。論文主要研究?jī)?nèi)容如下:1病毒感染小鼠后的臨床癥狀在2,000TCID50的LY-2015株P(guān)RV感染組,小鼠在病毒感染48h后開始出現(xiàn)被毛粗亂、食欲不振;在病毒感染60h后,小鼠表現(xiàn)出精神振奮,抓撓頭面部皮膚;病毒感染84h后,多數(shù)小鼠因瘙癢抓撓致皮膚破潰,小鼠死亡。用相同感染量的Halv株P(guān)RV滴鼻感染小鼠時(shí),未見任何臨床癥狀。在10,000TCID50的Halv株P(guān)RV感染組,小鼠在病毒感染72h后開始出現(xiàn)與LY-2015株P(guān)RV感染組相似的臨床癥狀;在病毒感染96h后,大量小鼠死亡。結(jié)果表明LY-2015株P(guān)RV毒力較Halv株P(guān)RV強(qiáng)。2病毒感染后的上行傳導(dǎo)通路LY-2015株P(guān)RV感染小鼠后,病毒首先出現(xiàn)在鼻黏膜下層細(xì)胞內(nèi),然后在三叉神經(jīng)相關(guān)神經(jīng)核團(tuán)、交感神經(jīng)相關(guān)神經(jīng)核團(tuán)、面神經(jīng)相關(guān)神經(jīng)核團(tuán)內(nèi)均檢測(cè)到PRV。表明變異病毒LY-2015株P(guān)RV滴鼻感染小鼠后可沿三叉神經(jīng)、交感神經(jīng)與面神經(jīng)通路上行傳導(dǎo),致中樞感染。Halv株P(guān)RV滴鼻感染小鼠后,在三叉神經(jīng)相關(guān)核團(tuán)、交感神經(jīng)相關(guān)核團(tuán)、副交感神經(jīng)相關(guān)核團(tuán)與面神經(jīng)相關(guān)核團(tuán)內(nèi)檢測(cè)到PRV。說明經(jīng)典病毒Halv株P(guān)RV滴鼻感染小鼠后可沿三叉神經(jīng)、交感神經(jīng)、副交感神經(jīng)和面神經(jīng)上行傳導(dǎo)。3病毒感染小鼠后在腦內(nèi)的分布2,000TCID50的LY-2015株P(guān)RV感染小鼠后,病毒多分布于脊髓,腦橋和延髓,大腦內(nèi)僅在丘腦可見少量病毒。在病毒感染84h后,小鼠腦內(nèi)的病毒分布最為廣泛。10,000TCID50的Halv株P(guān)RV感染組,病毒不僅分布于脊髓、腦橋和延髓,大腦中也存在多量病毒。病毒感染小鼠96h后在腦內(nèi)的分布最為廣泛。結(jié)果表明,經(jīng)典病毒Halv株P(guān)RV感染小鼠后沿軸突上行傳導(dǎo)較變異病毒LY-2015株P(guān)RV慢;且LY-2015株P(guān)RV的毒力較強(qiáng),小鼠在病毒感染后過早死亡,僅少量病毒傳導(dǎo)至中樞,分布于下丘腦外側(cè)部、丘腦下中間核腹側(cè)部等。此外,在2,000TCID50的LY-2015株P(guān)RV感染組,小鼠在病毒感染后120h,IHC染色呈PRV陰性,但I(xiàn)SH染色呈PRV陽(yáng)性,結(jié)果表明小鼠腦內(nèi)存在病毒基因,但病毒未復(fù)制。結(jié)果表明,感染PRV后,耐過小鼠呈隱性感染,病毒基因整合于小鼠外周神經(jīng)細(xì)胞核內(nèi),但不復(fù)制產(chǎn)生新的病毒病毒粒子。以上結(jié)果表明,LY-2015株P(guān)RV毒力強(qiáng)于Halv株P(guān)RV;Halv株P(guān)RV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時(shí),變異病毒LY-2015株P(guān)RV感染小鼠后沿三叉神經(jīng)、交感神經(jīng)與面神經(jīng)上行傳導(dǎo),但病毒粒子多分布于延髓和腦橋;經(jīng)典病毒Halv株P(guān)RV感染小鼠后可沿三叉神經(jīng)、交感神經(jīng)、副交感神經(jīng)與面神經(jīng)上行傳導(dǎo),在小鼠脊髓、腦橋、延髓和大腦中均存在多量的病毒分布。在有效感染中耐過小鼠呈潛伏性感染,不表現(xiàn)出任何臨床癥狀,病毒基因整合到小鼠外周神經(jīng)細(xì)胞核中,但不復(fù)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.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S852.65

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