| 牛布氏杆菌PCR检测方法的建立 |
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| 引用本文： | 王素华,王忠才,杜爱芳.牛布氏杆菌PCR检测方法的建立[J].中国畜牧兽医,2012,39(4):79-81. |
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| 作者姓名： | 王素华 王忠才 杜爱芳 |
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| 作者单位： | 1. 温州出入境检验检疫局,浙江温州 325027;2. 浙江大学动物预防医学研究所,浙江杭州 310029 |
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| 基金项目： | 浙江出入境检验检疫局立项项目(2011ZK019) |
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| 摘 要： | 根据GenBank上公布的牛布氏杆菌外膜蛋白OMP31的基因序列设计特异性引物,对牛布氏杆菌样品进行PCR扩增并将产物克隆到pMD18-T载体后测序。结果表明,扩增的目的片段大小为602 bp,与预期扩增序列同源性为99.7％;该PCR检测体系的特异性强,不能在非布氏杆菌 DNA中扩增出条带;敏感性高,最低检测的DNA含量为0.9 pg/μL。该检测体系的成功构建为牛布氏杆菌病的检测、鉴定和流行病学调查提供了有力的技术支持。
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| 关 键 词： | 牛 布氏杆菌 PCR 检测方法 |
| 收稿时间： | 2011-01-02 |
Establishment of PCR for Detecting Bovine Brucella |
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| WANG Su-hua
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WANG Zhong-cai
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DU Ai-fang.Establishment of PCR for Detecting Bovine Brucella[J].China Animal Husbandry & Veterinary Medicine,2012,39(4):79-81. |
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| Authors: | WANG Su-hua WANG Zhong-cai DU Ai-fang |
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| Institution: | 1. Wenzhou Entry-exit Inspection and Quarantine Bureau, Wenzhou 325027, China;2. Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029,China |
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| Abstract: | A pair of primers were designed according to the sequence of specific Brucella gene of Outer membrane protein(OMP31),the OMP31 gene was amplified by PCR and cloned into pMD18-T vector,and then the gene sequence was detected.The results showed that the target gene band at a length 602 bp was amplified by PCR,with a homology of 99.7% to the gene sequence reported in GenBank.The experiments had proved that PCR assay possessed a high specificity,DNA bands couldn’t amplified in other bacterial except Brucella.And the sensitivity test results indicated that PCR assay was more sensitive,which could detect OMP31 with only 0.9 pg/μL DNA.The successfully construction of the PCR assay provides strongly technical support for detection,identification and epidemiological investigations of bovine brucellosis. |
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| Keywords: | bovine Brucella PCR detection |
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