【作者】 王波；

【导师】 田夫林； 姜世金；

【作者基本信息】 山东农业大学 ， 预防兽医学， 2009， 硕士

【摘要】 猪瘟(Hog Cholera,HC或Classical Swine Fever,CSF)是由猪瘟病毒(HCV或CSFV)引起的一种高度接触性热性传染病。以出血和发热为主要特征,并引起妊娠母猪流产、胎儿畸变、慢性营养性消耗、淋巴细胞和血小板减少等免疫系统受损、继发或并发细菌或其他病毒感染等临床表现,呈急性或慢性经过,是一种对猪危害性极大的传染病,被国际兽疫局(OIE)列为必须上报的疾病。一直以来我国采用猪瘟疫苗免疫对猪瘟进行预防和控制,在降低猪瘟的感染率和发病率方面取得了很好的效果,但是近年来在实际生产中猪瘟仍时有发生,特别是仔猪的非典型猪瘟更为常见。仔猪发生猪瘟除了首免时间不当,母源抗体对免疫接种猪瘟疫苗的干扰作用造成的免疫失败之外,实际的免疫效果也是很重要的一个因素。同时,对猪瘟疫苗质量的评定,也是猪瘟免疫的重要前提。本研究共分为三个部分:1、采用阻断ELISA试验方法对不同日龄仔猪的猪瘟母源抗体进行系统检测,其结果表明:哺乳前仔猪抗体水平为0,第1d抗体水平达到最高,之后缓慢下降。30日龄平均抗体水平仍然在保护水平之上,因而过去常用的20日龄首免时间应该推迟;根据20日龄仔猪断乳时的抗体效价和30日龄的抗体效价推算,仔猪猪瘟母源抗体半衰期为6.8天。2、采用阻断ELISA试验方法检测三种不同猪瘟免疫程序对仔猪的免疫效果,免役程序分别为(0、35,60)、(0、60)和(35、60),其结果表明:对仔猪进行超前免疫(0、60)的免疫效果比其他免疫程序的免疫效果好。不同疫苗免疫的仔猪猪瘟抗体检测结果表明,仔猪平均抗体阻断率差异极显著(P<0.01),因而制定适宜的免疫程序,应该根据猪瘟疫苗质量而定。3、利用荧光定量RT-PCR和阻断ELISA试验方法分别对猪瘟疫苗的病毒含量和猪瘟疫苗中外源病毒(BVDV)的污染进行了检测。荧光定量RT-PCR对猪瘟疫苗病毒含量检测的结果表明:15种猪瘟疫苗中的病毒含量存在很大差异,从0.87×106～38.45×106病毒颗粒数/头份不等。此方法操作简单,速度快,定量准确,重复性好,可以在短时间内同时对大量样品进行定量检测,因而作为猪瘟疫苗效价的评定是可行的。但是荧光定量RT-PCR对操作人员的熟练程度要求严格;猪瘟疫苗中BVDV抗原检测的结果表明:3种脾淋苗中都没有BVDV抗原,而12种细胞苗中,有6种检测BVDV抗原阳性,污染率达50%。因此,猪瘟疫苗中外源病毒(BVDV)污染的检测,也是疫苗合格与否的重要指标。总之,对猪瘟疫苗质量的评定是猪瘟免疫的重要前提。更多还原

【Abstract】 Classical swine fever (Hog Cholera,HC or Classical Swine Fever,CSF) is a highly contagious disease in pigs caused by the Classical Swine Fever virus. The main characteristics of CSF is bleeding and hemorrhagic, and it caused abortion of pregnant sows, fetal aberration, chronic nutritional consumption, immune system injury, such as reduction of lymphocytes and thrombocytopenia, clinical manifestations secondary or concurrent such as bacterial infection or other virus infection, acute or chronic, and is a great danger of infectious diseases to pigs, the disease must be reported by the Office International des Epizooties (OIE).China has been the use of classical swine fever vaccine to prevent and control classical swine fever, and it achieves good results in reducing Classical swine fever incidence and infection. However, classical swine fever still occurs from time to time in recent years in the actual production, especially atypical Classical swine fever in piglets.In addition to the inappropriate first immune time of classical swine fever vaccine and interference of maternal antibodies of occurrence of classical swine fever in piglets,the practical effect of immunization is very important as a factor. At the same time, the assessment of the quality of classical swine fever vaccine is also an important prerequisite for classical swine fever immunization.This research consists of three parts as follows:PartⅠ: Blocking ELISA test methods is adopted to check out different day-old piglets of classical swine fever maternal antibodies. The result showed that the titers of the maternal antibodies in piglets were zero before suckle, arrived the highest level in the first day, and then slowly declined. The average 30-day-old antibodies level is still above the level of protection, Therefore the first immune time which commonly used in the past 20-day-old should be free to postpone. According to antibody titers of 20-day-old piglets weaned and antibody titers of 30-day-old piglets, half-life of Hog cholera maternal antibody in piglets is 6.8d.PartⅡ: Blocking ELISA test methods is adopted to check out immune effects of three different classical swine fever immunization procedure on piglets. Immune procedure are (0,35,60)、(0,60) and (35,60) respectively.The results of CSF antibodies by different vaccines immunization show that the average rate of piglets in differences blocking antibody are extremely significant (P < 0.01),so the development of appropriate immunization procedures should be based on the effect of classical swine fever vaccine.PartⅢ: Fluorescence quantitative RT-PCR and Blocking ELISA testing methods are adopted to detect the content of virus and exogenous virus (BVDV) for classical swine fever respectively. The evaluation results of titer of classical swine fever vaccine by fluorescence quantitative RT-PCR show that Virus content of 15 kinds of classical swine fever vaccine are very different, from 0.87×106 to 38.45×106 virus particles/mL. This method is simple、fast、accurate quantitative、good reproducibility, and a large number of samples can be quantitatively detected in a short time. Thus, it is feasible as titer assessment of classical swine fever vaccine. However, it demands operator proficiency strictly; The test results of BVDV antigen in Classical swine fever vaccine show that three kinds of spleen and lymph vaccine do not contain BVDV antigen, and there are six kinds of testing positive for BVDV antigen in twelve kinds of cell vaccine, contamination rate was 50%. Therefore, the detection of BVDV antigen in classical swine fever vaccine is an important indicator of vaccine whether qualified or not. In short, the quality of the assessment of classical swine fever vaccine is an important prerequisite for classical swine fever immunization.更多还原