牛支原体间接免疫酶标组织化学检测方法的建立  

Establishment of the Indirect Immuno-histochemical Method for Detecting Mycoplasma bovis

作　　者：金云云[1] 剡根强[1] 王静梅[1] 石刚[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832003

出　　处：《中国畜牧兽医》2013年第5期26-30,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家科技支撑计划课题(2012BAD43B02);石河子市农业科技攻关计划(2011NY10)

摘　　要：为建立检测牛支原体的间接免疫组织化学(简称免疫组化)方法,分别以鸡抗牛支原体(Mycoplasma bovis)IgY、羊抗鸡IgG-HRP为一抗和二抗,对牛支原体菌体涂片和牛支原体阳性肺脏组织切片进行免疫组化染色,并对染色条件进行优化,确定组织触片及组织切片的最佳免疫组化条件。在最佳条件下对病死犊牛病料切片进行免疫组化检测,并以细菌分离培养和PCR方法进行验证。结果显示,免疫组化法阳性标本与分离培养和PCR结果相符,而阴性对照和替代试验均为阴性。结果表明本试验建立的间接免疫组化方法可用于检测临床病例组织样本及培养物中的牛支原体,具有特异性和可靠性,为探究该病原在机体内的定位、动态分布规律及致病机理提供了手段。To establish indirect immuno-histochemical method for detection of Mycoplasma bovis（M.bovis）,the pure culture of M.bovis and the lung from the calve infected with the M.bovis were immuno-histochemically studied by using the chicken anti M.bovis IgY and goat anti-chicken IgG-HRP as the first and second antibody,and optimizing the staining conditions to determine the best immuno-histochemistry staining method.The method was used to detect the diseased sections of dead calves under optimal conditions,and the result was verified by isolation and PCR methods.The immuno-histochemistry positive specimens in line with isolation and PCR positive results,while the negative control and alternative test were negative.It showed that the indirect immuno-histochemical method established in this trial could be used to specifically and reliably detect M.bovis of tissue samples and culture in the clinical cases,it also could be provided to explore the site in the organism,the regularity of dynamic distribution and the mechanism of pathogenicity of this pathogen.

关 键 词：免疫组化 检测 牛支原体 

分 类 号：S858.23[农业科学—临床兽医学]