牛支原体肺炎流行RT-PCR方法的初步诊断  

Diagnosis of Mycoplasma Bovis Pneumonia by RT-PCR Method

作　　者：张永强[1] 张维[1] 王清华[1] 吴研功[1] 赵云玲[1] 赵永刚[1] 王志亮[1] 

机构地区：[1]中国动物卫生与流行病学中心国家外来动物疫病诊断中心,山东青岛266032

出　　处：《中国动物检疫》2011年第6期48-49,55,共3页China Animal Health Inspection

摘　　要：设计5对引物,对山东某一牛交易市场采集的64份鼻腔棉拭子样品进行了病原学检测。对所有样品进行差异脲原体、牛支原体、牛生殖道支原体、牛眼支原体和牛传染性胸膜肺炎(CBPP)PCR病原学检测,PCR产物连接T载体测序。PCR结果显示,牛支原体阳性样品7份,牛生殖道支原体、牛眼支原体和牛丝状支原体全部为阴性;测序结果显示,所测样品核酸序列与牛支原体和无乳支原体核酸序列都具有很高的同源性。对核酸序列进行遗传进化树分析表明,相对于无乳支原体而言,所测核酸序列与牛支原体物种具有更近的遗传距离。Five pairs of primers were designed to diagnose a novel infectious respiratory disease in cattle in a market in Shandong Province. Including general primer for most species of Mycobacteriurn genus, using the other several pairs of primers, Ureaplasma diversum, M.bovis, M.bovigenitalium, M.bovoculi and CBPP gene segment could be amplified respectively by RT-PCR. All the 64 nasal cotton swab samples were detected by RT-PCR with general primers and 12 were positive. All the 12 samples were tested with the other five pairs of primers, 7 were M.bovis positive and Ureaplasma diversum, M.bovigenitalium, M.bovoculi and CBPP negative, other 5 samples were M.bovis, Ureaplasma diversum, M.bovigenitalium, M.bovoculi and CBPP negative. The amplified segment were inserted to pMDI8 plasmid and sequenced. The sequence BLAST showed that it enjoyed high linear sequence homology with M.bovis and M. agalactiae together. The phylogenetic tree showed that the amplified sequence had closer relationship to M.bovis than M.agalactiae.

关 键 词：牛支原体 病原学检测 遗传进化树 

分 类 号：S852.62[农业科学—基础兽医学] S858.23[农业科学—兽医学]