应用巢式PCR方法检测牛支原体肺炎  

A nested PCR method for detecting mycoplasma bovis pneumonia

作　　者：李大伟[1,2] 张彦明[2] 黄灿平[2] 谢建华[3] 熊仲良[3] 冉智光[3] 关平原[1] 范伟兴[2] 

机构地区：[1]内蒙古农业大学,内蒙古呼和浩特011118 [2]中国动物卫生与流行病学中心,山东青岛266032 [3]重庆市动物疫病预防控制中心,重庆渝北401100

出　　处：《中国动物检疫》2010年第4期28-29,共2页China Animal Health Inspection

摘　　要：根据Genbank发表的牛支原体全基因序列,设计2对特异性引物,建立了诊断牛支原体肺炎的巢式PCR方法,第一轮引物P1、P2扩增片段长度为1912bp,第二轮引物P3、P4扩增片段长度为422bp:该方法特异性试验未扩增出丝状支原体丝状亚种小克隆和无乳支原体特异性条带;敏感性试验证明扩增DNA最低含量达到10-7μg/mL,是单一PCR的103倍;用该方法对阳性病料进行检测,均扩增出特异性片段。该体系的成功构建,可以为牛支原体肺炎的活体检测,和流行病学调查提供技术支持。Based on the reports and sequences published in Genbank, two pairs of specific primers were designed according to the conservative compass. The primers were designed for the establishment of the nested PCR. The amplificated product of primer P1 and P2 was about 1912bp, and the amplificated product of primer P3 and P4 was about 422bp. The specific tests showed that no fragment while detecting Mycoplasma mycoides subsp.mycoides small Colony Type and Mycoplasma agalaetiae. Results of sensitivity showed the detection limit of this method was 10-Tug/ml. Au positive samples were amplified with this method. This specific and sensitive method is suitable for the detection of living body Mycoplsma bovis pneumonia and gived proofs for the research of epidemiological analysis.

关 键 词：牛支原体 巢式PCR 活体检测 

分 类 号：S852.62[农业科学—基础兽医学]