猪肺炎支原体斑点杂交检测方法的建立及应用

为建立猪肺炎支原体斑点杂交检测方法,并用于猪肺样品的检测,根据GenBank中登录的猪肺炎支原体(Mhp)P36基因序列,设计并合成一条地高辛标记的DNA寡核苷酸探针。进行Mhp斑点杂交反应,建立最佳反应体系和反应条件,对该方法进行特异性和敏感性试验,并采用最佳反应体系和反应条件在尼龙膜上对猪肺组织样品进行检测。结果表明,该检测方法可有效从可疑病料的猪肺组织中检出Mhp感染,检出率为39.5%。本研究建立的Mhp斑点杂交检测方法快速、敏感、特异,对Mhp的临床检测具有较好的应用价值。

Establishment and Application of the Dot Blot Hybridization Method for Detecting Mycoplasma hyopneumonia

The aim of this paper was to establish dot blot hybridization method to detect Mycoplasma hyopneumoniae(Mhp),which would be used to detect the suspect swine lung samples.According to the sequence of Mhp P36 gene,a digoxin-labbled DNA probe was designed and synthesized.The optimum dot blot hybridization reaction system and conditions were established.The sensitivity and specificity tests were done,and then the lung samples of swine were detected by the established system and conditions on nylon membrane.The dot blot hybridization detection method of Mhp was established,the suspect swine infected with Mhp could be detected effectively by this method,and the detection rate was 39.5%.The established Mhp dot blot hybridization detection method was rapid,sensitive and specific.Moreover,it would be helpful for clinical detection of Mhp.