绵羊肺炎支原体免疫组织化学及间接免疫荧光检测方法的建立

Establishment of indirect immunofluorescence assay and immunohistochemistry assay for detection of Mycoplasma ovipneumonia

以病理组织学观察为基础,建立检测绵羊肺炎支原体的间接免疫组织化学和间接免疫荧光方法,以绵羊肺炎支原体标准血清为一抗,分别以家兔抗山羊Ig G-HRP和家兔抗山羊Ig G-FITC为二抗,对绵羊肺炎支原体阳性肺组织切片及阴性对照组切片进行HE染色、免疫组织化学染色及间接免疫荧光染色,并优化染色条件,以确保组织切片在最佳的条件下进行鉴定。结果显示,本试验所建立的免疫组织化学方法与PCR方法对病样的检出率相符,且高于传统的分离培养法。结果表明,本试验建立的间接免疫组织化学方法和间接免疫荧光方法可用于对临床病例的组织样本检测,具有很强的特异性和可靠性,可直观地观察和探究该病原在机体内的定位、动态分布以及致病机理,以期为临床诊断提供更可靠的方法。

Indirect immunohistochemistry staining（IIS） detection method of Mycoplasma ovipneumonia was developed based on histopathological observations in this study.The standard serum of M.ovipneumoniae was treated as a primary antibody,while the rabbit anti-goat Ig G-HRP and rabbit anti-goat Ig G-FITC were treated as a secondary antibody.Then sections of the sheep lungs infected with Mycoplasma pneumoniae and lungs in the negative control group were treated with HE staining,immunohistochemistry staining and immunofluorescence staining,and the staining conditions were optimized.As the results showed,the relevance rate of IIS detection method which was established in our experiment was consistent with the PCR method,and its sensibility was much higher than that of the traditional isolation method.IIS detection method and indirect immunofluorescence assay could be applied for detection of clinical samples with high specificity and accuracy.The results suggest that it is a reliable detection method for clinical diagnosis.