内蒙古地区乳样中牛支原体的分离及PCR鉴定  

Isolation and PCR Identification of Mycoplasma bovis from Milk in Inner Mongolia

作　　者：许金朋 黄海碧[1] 王晓晖[1] 郝永清[1] 

机构地区：[1]内蒙古农业大学兽医学院微生物学与免疫学实验室,内蒙古呼和浩特010018

出　　处：《动物医学进展》2015年第4期24-27,共4页Progress In Veterinary Medicine

基　　金：国家科技支撑计划项目(2012BAD12B09-02;2012GB2A400058)

摘　　要：为检测内蒙古地区某牛场患乳房炎的奶牛乳样中是否存在牛支原体,同时建立直接提取乳样中牛支原体DNA进行PCR检测的方法,本研究无菌采集乳房炎乳样,通过支原体的分离培养、形态学观察和生化试验,初步鉴定分离株为牛支原体,然后提取液体培养基菌体DNA进行牛支原体特异性PCR鉴定,同时,采用Chelex-100法直接提取原乳样中菌体DNA进行PCR鉴定,并测序分析扩增片段序列。液体培养物提取DNA与Chlex-100法直接提取原乳样菌体DNA进行特异性PCR均扩增出目的条带,该片段序列与GenBank中牛支原体oppD/oppF基因的同源性达到99.8%,证实分离株为牛支原体。说明Chlex-100法可直接提取乳样中牛支原体DNA进行快速PCR鉴定。To detect the presence of Mycoplasma bovis in milk samples from cows in a dairy farm in Inner Mongolia suffering from mastitis,while establishing extraction method of mycoplasma DNA directly from milk samples for PCR detection,preliminary identification of Mycoplasma bovis from Inner Mongolia mastitis milk samples was carried out by culture,morphological characters and biochemical tests.Then DNA was extracted from broth cultures and milk samples by Chelex 100 method for specific PCR tests.Finally,the sequence of the PCR products was analyzed.Expected PCR products were both amplified when the DNA was extracted from broth cultures and milk samples.The sequece of the PCR products had 99.8%identity with corresponding sequence of oppD/oppF gene in GenBank.In conclusion,Mycoplasma bovis was isolated from bovine mastitis milk samples in Inner Mongolia,and a simple extraction of Mycoplasma bovis DNA for PCR using chelex 100 from milk samples was built.

关 键 词：牛支原体 乳房炎 牛乳 Chelex-100法 PCR鉴定 

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