牛支原体病PCR检测方法的建立及初步应用

Establishment and Preliminary Application of PCR Assay for the Detection of Mycoplasma bovina

为快速确诊肉牛牛支原体病,根据牛支原体 OPPD/F 基因序列设计1对引物,通过反应条件的优化,建立牛支原体PCR诊断方法。结果表明:该方法可从牛肺、鼻拭子临床样本和其临床样本的培养物中直接检测到牛支原体病病原核酸,该方法可检测的病原核酸最小浓度约为1×10^-7 μg,整个检测过程仅需3.5 h。该方法具有快速、灵敏性高、特异性强等特点,对牛支原体感染的诊断、防治与预后评估具有重要意义。

To rapidly diagnose Mycoplasma bovina in beef cattle,a pair of primers were designed according to the OPPD/F gene sequence of M.bovina ,and the PCR diagnosis method of M.bovina was established by optimizing the reaction conditions.Results:The method could detect the pathogenic nucleic acid of M.bovina directly from the clinical samples of bovine lung and nose swabs and the culture of the clinical samples.Minimum detectable concentration was 1×10^-7 μg,and the detecting time was 3.5 h.It was concluded that the established PCR assay was rapid,highly specific and sensitive,which could be important in disease diagnosis,control applications and prognostic estimation of M.bovina.