羊支原体肺炎胶体金免疫层析检测方法的建立及应用  

Establishment and Application of Colloidal Gold Immunochromatographic Assay for Detecting the Mycoplasmal Pneumonia of Sheep and Goats

作　　者：王璇 潘淑惠 吴玙彤 黄波 刘显明 宫伟 文正常 WANG Xuan;PAN Shu-hui;WU Yu-tong;HUANG Bo;LIU Xian-ming;GONG Wei;WEN Zheng-chang(Institute of Animal Husbandry and Veterinary Medicine of Guizhou Academy of Agricultural Sciences,Guiyang 550005,China;Animal Epidemic Prevention and Control Center of Weining County of Guizhou Province,Weining 553100,China;Animal and Poultry Breed Improvement Station of Weining County of Guizhou Province,Weining 553100,China)

机构地区：[1]贵州省农业科学院畜牧兽医研究所,贵州贵阳550005 [2]贵州省威宁县动物疫病预防控制中心,贵州威宁553100 [3]贵州省威宁县畜禽品种改良站,贵州威宁553100

出　　处：《中国兽医杂志》2020年第10期38-41,45,共5页Chinese Journal of Veterinary Medicine

基　　金：贵州省科技计划项目[黔科合NZ(2013)3025]。

摘　　要：本试验应用绵羊支原体肺炎RPS 11基因重组蛋白为免疫抗原,制备鼠RPS11单克隆抗体,经间接ELISA特异性抗体效价检测,小鼠腹水单克隆抗体的效价均高于105。将制备的鼠RPS11单抗进行胶体金标记、纯化、质量鉴定及条件优化,建立羊支原体肺炎胶体金免疫层析检测方法。应用制备的免疫金标诊断试纸条对临床180份疑似血液样本进行检测,并与ELISA检测结果进行比较,结果显示,2种方法的阳性符合率为91.11%;特异性试验结果显示,试验诊断检测试纸条不与羊其他常见感染病原发生交叉特异性反应。因此,研制的金标诊断检测试纸条为羊支原体肺炎的早期诊断检测及流行病学调查奠定了良好的技术基础。In this study,the recombinant plasmid pET28a-RPS11 of Mycoplasma ovipneumoniae RPS 11 gene was used as immune antigen.The mouse RPS11 monoclonal antibody was prepared.The specific antibody titer of indirect ELISA was detected and the effect of the monoclonal antibody was determined in ascites of mice.A colloidal gold immunochromatographic assay for detection of Mycoplasmal pneumonia of sheep and goats was established by labeling,purifying,quality identification and optimizing the conditions of the prepared mouse RPS11 monoclonal antibody.The immune gold label diagnostic strip was used to detect 180 suspected blood samples and the result was compared with that by the ELISA.The results showed that the positive coincidence rate with ELISA was 91.11%.The specificity test showed that the test strip did not cross-react with other common infectious pathogens in sheep and goats.Therefore,the developed gold-label diagnostic test strip lays a good technical foundation for the early diagnosis and detection of Mycoplasmal pneumonia of sheep and goats and epidemiological investigation.

关 键 词：羊支原体肺炎 单克隆抗体 胶体金免疫层析 

分 类 号：R375.2[医药卫生—病原生物学]