鉴别牛羊布鲁菌实时荧光定量PCR方法的建立  

Establishment of Real-time Fluorescence Quantitative PCR to Identify B. abortus and B. melitensis

作　　者：苏娇[1] 王艳杰[1] 智达夫[1] 关平原[1,2] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]农业部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010018

出　　处：《畜牧与饲料科学》2013年第5期7-12,共6页Animal Husbandry and Feed Science

摘　　要：[目的]建立准确、快速检测布鲁菌、羊种布鲁菌、牛种布鲁菌的方法。[方法]以BCSP31作为布鲁菌菌属特异性基因,以IS711插入元件作为布鲁菌菌种特异性标志,设计引物和Taqman探针;分别构建标准阳性质粒模板,将其10倍倍比稀释作多重实时荧光定量PCR标准曲线,评价多重实时荧光定量PCR反应体系的敏感性、特异性、重复性。[结果]建立了鉴别牛种、羊种布鲁菌的实时荧光定量PCR方法,并用该方法对临床样品进行检测。[结论]所建立的多重实时荧光定量PCR诊断方法能够快速、准确地鉴别牛种和羊种布鲁菌。[Objective] The aim was to establish an accurate and rapid diagnostic method for detection of Brucella spp. ,Brucella abortus and Brucella melitensis. [Method] The primers and Taqman probe were designed by using BCSP31 as the specific gene and an ISTll element as the specific insertion for Brucella spp. identification. Standard positive plasmid templates were constructed respectively, and its ten-fold serial dilutions were used for making multiple quantitative PCR standard curve to evaluate the sensitivity, specificity and reproducibility of the multiplex real-time fluorescence quantitative PCR reaction system. [Result] The real-time fluorescence quantitative PCR for the identification of B. abortus and B. melitensis was established and used for the testing of clinical samples. [ Conclusion ] The establishment of the multiplex real-time fluorescence quantitative PCR diagnostic method could identify B. abortus and B. melitensis quickly and accurately.

关 键 词：实时荧光定量PCR 布鲁菌 牛种布鲁菌 羊种布鲁菌 TAQMAN探针 

分 类 号：S854.43[农业科学—临床兽医学]