【摘要】：近年來，乙型腦炎病毒（JEV）在我國(guó)甚至亞洲地區(qū)廣泛流行，威脅著人和家畜的安全，經(jīng)過這些年我國(guó)政府及相關(guān)部門的努力，患乙型腦炎的人的發(fā)病率及死亡率不斷下降，但是對(duì)于豬的乙型腦炎的預(yù)防還沒有豬專用的乙型腦炎病毒疫苗，目前一般采用弱毒疫苗SA14-14-2對(duì)豬進(jìn)行免疫。因此，根據(jù)當(dāng)前我國(guó)養(yǎng)豬行業(yè)的形勢(shì)，需要一種豬專用的乙型腦炎病毒疫苗，本實(shí)驗(yàn)室研制的含乙型腦炎E抗原基因重組痘苗病毒疫苗rVTT-JEVE具有良好的免疫效果，但是目前尚不能證明該疫苗的安全性。 本研究對(duì)本實(shí)驗(yàn)室研制的重組痘苗病毒疫苗rVTT-JEVE的安全性進(jìn)行評(píng)價(jià)，首先對(duì)重組痘苗病毒rVTT-JEVE的理化特性進(jìn)行研究，通過將其進(jìn)行溫度、酸堿度、乙醚、胰蛋白酶、氯仿、超聲波及紫外線等理化因子的適當(dāng)?shù)牟煌兞康奶幚�，檢測(cè)其經(jīng)處理后的病毒滴度的變化，證明其對(duì)高溫敏感，對(duì)酸堿有一定的耐受性，對(duì)氯仿、胰蛋白酶、超聲波、紫外線均有一定程度的敏感性，完成了理化特性的研究。 首先應(yīng)用小鼠進(jìn)行了免疫相關(guān)研究，且對(duì)重組痘苗病毒疫苗株rVTT-JEVE的安全性進(jìn)行了初步評(píng)價(jià)，免疫小鼠后產(chǎn)生的特異性抗體水平顯著高于弱毒疫苗組和對(duì)照組（P0.05），經(jīng)RT-qPCR方法檢測(cè)重組痘苗病毒在小鼠體內(nèi)殘留發(fā)現(xiàn)，重組痘苗病毒在小鼠體內(nèi)5-7d后拷貝數(shù)明顯減少，通過分析其體液免疫及病毒殘留，證明該候選疫苗具有良好的體液免疫且病毒在小鼠體內(nèi)殘留僅5-7d。通過豬體免疫，分析其免疫特性、病毒殘留和環(huán)境釋放，，證實(shí)了該重組痘苗病毒候選疫苗株rVTT-JEVE有良好的免疫特性，重組痘苗病毒在豬體內(nèi)殘留僅3d且不會(huì)向環(huán)境中釋放，表明該候選疫苗具有良好安全性。
[Abstract]:In recent years, Japanese encephalitis virus (JEV) has been widely prevalent in China and even in Asia, threatening the safety of human beings and livestock. Through the efforts of our government and related departments, the morbidity and mortality of people suffering from encephalitis B have been decreasing. However, for the prevention of swine encephalitis B, there is no swine specific vaccine against Japanese encephalitis virus. Currently, attenuated vaccine SA14-14-2 is generally used to immunize pigs. Therefore, according to the current situation of pig industry in our country, we need a kind of special type B encephalitis virus vaccine. The recombinant vaccinia virus vaccine rVTT-JEVE containing E antigen gene of Japanese encephalitis developed by our laboratory has good immune effect. But it is not yet possible to prove the vaccine's safety. In this study, the safety of recombinant vaccinia virus vaccine rVTT-JEVE was evaluated. Firstly, the physical and chemical properties of recombinant vaccinia virus rVTT-JEVE were studied. The proper treatment of different variables of physical and chemical factors, such as ultrasonic wave and ultraviolet ray, to detect the changes of virus titer after treatment, proved that it is sensitive to high temperature, has certain tolerance to acid and alkali, and is resistant to chloroform, trypsin, ultrasonic wave, and so on. UV rays were sensitive to a certain degree and the physicochemical properties were studied. First of all, immune-related studies were carried out in mice, and the safety of recombinant vaccinia virus vaccine strain rVTT-JEVE was preliminarily evaluated. The specific antibody level of immunized mice was significantly higher than that of attenuated vaccine group and control group (P0.05). The recombinant vaccinia virus remained in mice by RT-qPCR, and the copy number of recombinant vaccinia virus in mice was significantly decreased after 5-7 days. By analyzing its humoral immunity and virus residues, it was proved that the candidate vaccine had good humoral immunity and the virus remained only 5-7 days in mice. The immune characteristics, virus residues and environmental release of the recombinant vaccinia virus candidate strain rVTT-JEVE were analyzed. The recombinant vaccinia virus remained in pigs for only 3 days and could not be released into the environment, the results showed that the recombinant vaccinia virus candidate strain rVTT-JEVE had good immune properties. The result shows that the candidate vaccine is safe.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S859.79

【共引文獻(xiàn)】

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5 王

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