【作者】 刘中奇；

【导师】 熊焰；

【作者基本信息】 四川农业大学 ， 预防兽医学， 2005， 硕士

【摘要】 本研究从猫瘟热病毒、猫鼻气管炎病毒、猫鼻结膜炎病毒三联弱毒活疫苗中分离猫瘟热病毒弱毒株,通过对分离株进行病毒理化特性检测、双向琼脂扩散试验检测和病毒粒子形态电镜观察,最终确认从三联弱毒疫苗中分离到猫瘟热病毒,并以此分离株作为诊断抗原。阳性抗体用国产单联猫瘟热灭活疫苗免疫试验动物制备。通过对酶联免疫吸附试验(ELISA)的操作技术加以适当改进,使酶标板的吸附作用提高了80%以上,进而使临床用ELISA方法检测FPV的灵敏度得以大大提高。根据相关资料提供的方法,对HA/HI的操作方法做了改进,建立了一套适合用于临床检测FPV的HA/HI方法。检测结果显示,ELISA和HA/HI的吻合率达到93%以上。将ELISA和HA/HI两种血清学方法相结合,用于野生动物FPV的诊断,不仅克服了国内以往主要依靠流行病学、临床症状和病理变化等进行诊断的弊端;而且显著提高了临床检验的敏感性、特异性和可操作性。 以FPV分离株为材料免疫试验动物,检测其分离毒株的免疫原性。结果显示FPV具有良好的免疫原性,能够诱导机体产生坚强的体液免疫和细胞免疫。从不同种类疫苗免疫金钱豹后的结果可以看出,经一次免疫后弱毒活疫苗在第36w、灭活疫苗(单联或三联)在第23w的ELISA检测P/N仍可达到2以上,这为野生动物制定免疫程序提供了可参考的理论依据。更多还原

【Abstract】 The study was aided by the Ailuropoda Fund and designed on account of the Chengdu zoo’ s topic that the Detection of the feline distemper’ s serum antibody and the foundation of its immune programme about the grow wild feline animal such as Panthera pardus L, et al. The study regarded the attenuated feline panleukopenia virus as the diagnostic antigen which was isolated from the tri-combinant vaccine of Feline rhinotracheitis virus, calicivirus and panleukopenia virus . According to the tesults of Virus Physical and Chemical Character Detection, Agar Diffusion Reaction and Virion Shape Electron Microscope Observation, it was affirmed that the virion which was isolated from the tri-combinant vaccine was feline panleukopenia virus . Positive diagonistic antibody was made from the experimental animal which was immunized by single inactive FPV vaccine. The adsorption of the enzyme-labelled plate had been elevated about 80% by improveing the technique of ELISA. Furthermore,the sensitivity of ELISA had been elevated a lot when it was used to detecte FPV on clinic. Based on the related datum, we improved the techinique of HA/HI and established a suit of method which fitted the need for using on clinic. The results showed that the coincidence rate reached above 93% between ELISA and HA/HI. If ELISA and HA/HI were combined together when diagnosed FPV about the grow wild feline animal, it would not only overcome the shortcoming that the diagnosis of FPV was mainly depended on the epidemiology, the clinical symptom and the pathological change, but also elevate the sensitivity , the specificity and the facility of the FPV’s clinical diagnosis.In order to detect the immunogenicity of the isolated strain, we injected it to the experimental animal.The results showed that it possessed favourable immunogenicity. It could induce forcefulness humoral immunity and cell immunity. The results that the Panthera pardus L. were immunized by the variety classes FPV vaccine showed that the P/N of ELISA was above 2 at the 36th week after the attenuated FPV vaccine was immunized and at the 23th week after the inactivated FPV vaccine was immunized. The datum would provide referenceable theory evidence when the people institute the immune programme for the grow wild feline animal.更多还原