课题名称 新孢子虫病快速检测试剂盒的研制 Development of rapidly diagnostic kit of Neospora caninum 执行时间 2009年 1 月-2011年12 月 课题编号 200811110 项目来源 自治区高新技术项目办 资助经费 30 万 主 持 人 巴音查汗 结 题 否 是 成 员 岳成、季新成、王莲芳、刘建华、赛福鼎、张晓红、王常汉、王振宝、杨帆 摘 要 新孢子虫病(Neosporosis)是由犬新孢子虫引起的一种全球性的原虫病,该病对牛的危害最为严重,是世界范围内造成奶牛流产、产弱胎、死胎、木乃伊胎的主要原因之一,可垂直传播。新疆是我国重要的畜牧业生产基地,养牛业又是我区的主要支柱产业,新孢子虫病是危害畜牧业特别是奶牛业的一种重要寄生虫病。目前,国内尚无控制该病的有效办法,因此,研究高效、特异、敏感的诊断方法尤为重要,研制快速、准确度高、重复性好的诊断试剂盒已成为当前研究该病的重点。我国虽有用ELISA进行牛新孢子虫血清学诊断及流行学调查的报道,但均采用了国外生产的诊断试剂盒,价格昂贵,所采样本少,容易漏检,局限性很大,没深入探讨其流行病学规律且没有表现出感染率、感染强度。近年来在新孢子虫分子生物学研究中发现的P43(NcSRS2t)和P36(NcSAG1t)是新孢子虫的主要表面蛋白,是最有希望的新孢子虫病诊断抗原之一。因此,以 NcSAG1t/NcSRS基因编码的重组蛋白作为抗原,建立rELISA,诊断新孢子虫病是研究的焦点。 我们通过本项目建立rELISA检测方法和研制rELISA检测试剂盒,为国内新孢子虫病高效快速检测及日常监测工作的开展提供有效的技术保障,对于新疆兽医防疫体系建设的进步完善和动物疫病的防治具有重要意义。 Abstract Neosporosis is a kind of worldwide protozoal disease caused by Neospora caninum, The disease is most serious harm to cattle,is one of the main that causes the cows abortion,weak fetus,stillbirth and mummy fetus .It can vertical transmission. As an important production base of animal husbandry in Xinjiang autonomous region of China.Cattle industry is the major pillar industry of Xinjiang. Neosporosis is harmful to animal husbandry particular dairy industry is an important parasitic disease. Currently, there is no effective drug and vaccines for control of Neosporosis. Therefore, the study of efficient, specific and sensitive diagnostic method is particularly important. The development of rapid, accurate and reproducible diagnostic kits have become the focus of current research of the disease. Although performed by ELISA of Neospora serological diagnosis and epidemiological survey reported, but used the foreign production diagnostic kit. The foreign production diagnostic kit expensive, a small number of test samples, easily missed, great limitations. No further investigate the epidemiology of law and showed no infection rate, infection intensity. Recent in years in the study of molecular biology Neospora that discovered P43 (NcSRS2t) and P36 (NcSAG1t) is the major surface protein of Neospora. This surface protein is the most promising one diagnostic antigen in Neosporosis. Therefore, NcSAG1t/NcSRS gene encoding the recombinant protein as an antigen, establish rELISA, diagnosis neosporosis is the focus of the study. Through this project,we established rELISA detection methods and developed rELISA test kits, that for domestic Neosporosis fast and efficient testing and routine monitoring to conduct provide effective technical support,for veterinary epidemic prevention system further improve and the prevention and control of animal diseases has great significance.