牛支原体的分离鉴定及16S rRNA基因序列分析  

Isolation,Identification and Sequence Analysis of 16 S rRNA Gene of Mycoplasma bovis

作　　者：伍晓红[1,2] 储岳峰[1] 张轩[1] 赵萍[1] 高鹏程[1] 贺英[1] 逯忠新[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部草食动物疫病重点开放实验室,甘肃兰州730046 [2]西藏昌都左贡县兽医站,西藏左贡854400

出　　处：《动物医学进展》2012年第12期35-37,共3页Progress In Veterinary Medicine

基　　金：农业部"西部之光"计划项目;国家科技基础性工作专项计划项目(2008FY210200);甘肃省自然科学基金项目(1107RJZA107)

摘　　要：为明确甘肃某牛场发生的疑似犊牛支原体肺炎的病原,利用支原体培养基从2份患肺炎的犊牛肺脏病料中分离到2个分离株,分别命名为GT-01和GT-02。通过菌落形态观察、生化试验、特异性PCR鉴定和16SrRNA测序比对对分离株进行鉴定。结果2个分离株在固体培养基上生长的菌落呈典型的"油煎蛋状",不能水解葡萄糖、甘露醇、精氨酸,不消化酪蛋白,PCR能扩增出牛支原体特异的1 911bp目的片段,16SrRNA基因序列与牛支原体国际标准株PG45的同源性分别为99.4%和99.6%。结果表明,2个分离株属于牛支原体。To identify the pathogen from calves showed pneumonia symptoms occurred in a cattle farm in Gansu province, two isolates （designated as GT-01 and GT-02） were isolated from two pneumonic lung tissue samples of calves using mycoplasma medium. Both isolates were characterized by the morphologic observation, biochemistry test, specific PCR test and sequence analysis of the 16 S rRNA gene. Colonies of two isolates grown on solid medi- um were typical fried-egg shape. Both isolates were unable to hydrolyze glucose, mannitol, arginine, and to digest casein. An expected gene fragment of 1 911 bp specific for Mycoplasma boris were amplified by PCR. Sequence a- nalysis of the 16 S rRNA gene revealed that GT-01 had an identity of 99.4 % and GT-02 had 99.6% identity with reference strain of PG45, respectively. The results indicated that the both isolates belonged to Mycoplasrna boris, which suggested that Mycoplasma boris infection has been prevalent in some areas of Gansu province.

关 键 词：牛支原体 分离 鉴定 16 S RRNA 

分 类 号：S852.62[农业科学—基础兽医学]