中国兽医科学2023，Vol.53Issue(4)：439-447,9.DOI:10.16656/j.issn.1673-4696.2023.0072
绵羊肺炎支原体特异性诊断靶标筛选及其间接ELISA方法的建立
Screening of specific diagnostic targets of Mycoplasma ovipneumoniae and establishment of the indirect ELISA
摘要
Abstract
The purpose of this study was to screen the specific antigen target of Mycoplasma ovipneumoniae (Mo) and establish an indirect ELISA method. The antigen target of Mo was screened, expressed and purified by Pull-down experiment combined with LC-MS analysis, and the reactivity of the purified protein was analyzed by Westernblot test. Then immunized mice, analyze the immunogenicity of the protein, prepare immune serum, establish an indirect ELISA method based on the target protein, and evaluate its performance. The results showed that the selected Mo pdhD protein was about 71 ku, which could still react with Mo positive sera diluted 1:4 000. The serum titer could reach 1:102 400. The established indirect ELISA method did not cross-react with positive serum such as Sheep pox case, and the coefficients of variation were within 12%, and the compliance rate with IHA method reached 82.5%. The above results showed that Mo pdhD protein has good reactivity and immunogenicity, and the established indirect ELISA method has good performance that can be used for serological diagnosis of Mo, which lays a foundation for further development of Mo ELISA antibody detection kit.关键词
绵羊肺炎支原体/Mo pdhD/免疫原性/间接ELISAKey words
Mycoplasma ovipneumoniae/Mo PdhD/immunogenicity/indirect ELISA分类
农业科技引用本文复制引用
李倩倩,储岳峰,高鹏程,葛家振,吴晓妮,田彤彤,许健,郝华芳,郑福英,蔺国珍..绵羊肺炎支原体特异性诊断靶标筛选及其间接ELISA方法的建立[J].中国兽医科学,2023,53(4):439-447,9.基金项目
国家重点研发计划项目(2022YFD1800704) （2022YFD1800704）
中央高校基本科研业务费专项资金项目(31920220137) （31920220137）
