| 布鲁氏菌BP26基因标记疫苗株的构建及鉴别PCR方法的建立 |
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| 引用本文： | 汪舟佳,甄清,乔凤,王玉飞,杜昕颖,钟志军,赵瑾,于雅琴,黄留玉,孙岩松,陈泽良.布鲁氏菌BP26基因标记疫苗株的构建及鉴别PCR方法的建立[J].微生物学报,2009,49(3):405-409. |
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| 作者姓名： | 汪舟佳 甄清 乔凤 王玉飞 杜昕颖 钟志军 赵瑾 于雅琴 黄留玉 孙岩松 陈泽良 |
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| 作者单位： | 1. 军事医学科学院疾病预防控制研究所,北京,100071
2. 吉林大学公共卫生学院,教育部重点实验室,长春,130021 |
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| 基金项目： | 国家自然科学基金，国家高技术研究发展计划(863计划) |
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| 摘 要： | 目的]由于现有的减毒活疫苗仍存在较强的毒力,因抗原与毒株的差异不大而很难区分疫苗免疫和自然感染等缺点,限制了现有布鲁氏菌减毒活疫苗的广泛应用.本文拟对布鲁氏菌的减毒活疫苗株M5进行遗传改造,克服这些缺点.方法]本研究利用同源重组的方法,用卡那抗性基因替换了布鲁氏菌减毒疫苗株M5的BP26基因,得到了新的标记疫苗株M5△BP26.分别用标记疫苗株和野生株侵染巨噬细胞和感染小鼠,比较分析标记株在细胞内和小鼠体内的存活能力.根据种特异性保守基因dnaK和缺失的BP26基因设计引物,建立双重PCR,用于区分标记株与野生株.结果]成功构建了.BP26基因标记疫苗株,细胞实验和动物实验结果表明,标记株仍能在胞内和小鼠内存活,具备作为减毒活疫苗的特性.小鼠实验结果显示,感染后两周野生株的细菌数为1022.9 ,而突变株为101.1 (P<0.01),至第3周野生株的细菌数为102.2 ,而突变株未能检出,表明与原疫苗株相比,标记株的感染力进一步减弱.根据DNA序列的差异,建立了能够区分标记疫苗株与野生株的双重PCR方法,标记株因只能扩增出一条带而能与野生株和毒株相区分,从而可以区分自然感染和疫苗免疫.结论]基因标记疫苗株的构建及鉴别PCR方法的建立,为布鲁氏菌疫苗的进一步研发奠定了基础.
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| 关 键 词： | 布鲁氏菌 26 kDa外膜蛋白 标记疫苗株 |
| 收稿时间： | 8/2/2008 12:00:00 AM |
| 修稿时间： | 2008/12/15 0:00:00 |
Construction of BP26 Tagged Vaccine Strain and Development of Discriminating PCR for Brucella |
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| Zhoujia Wang,Qing Zhen,Feng Qiao,Yufei Wang,Xinying Du,Zhijun Zhong,Jin Zhao,Yaqin Yu,Liuyu Huang,Yansong Sun and Zeliang Chen.Construction of BP26 Tagged Vaccine Strain and Development of Discriminating PCR for Brucella[J].Acta Microbiologica Sinica,2009,49(3):405-409. |
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| Authors: | Zhoujia Wang Qing Zhen Feng Qiao Yufei Wang Xinying Du Zhijun Zhong Jin Zhao Yaqin Yu Liuyu Huang Yansong Sun and Zeliang Chen |
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| Institution: | Institute of Disease Control and Prevention, Academy of Military Medical Science, Beijing 100071, China;School of Public Health, Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130021, China;School of Public Health, Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130021, China;Institute of Disease Control and Prevention, Academy of Military Medical Science, Beijing 100071, China;Institute of Disease Control and Prevention, Academy of Military Medical Science, Beijing 100071, China;Institute of Disease Control and Prevention, Academy of Military Medical Science, Beijing 100071, China;School of Public Health, Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130021, China;School of Public Health, Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130021, China;School of Public Health, Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130021, China;School of Public Health, Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130021, China;School of Public Health, Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130021, China |
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| Abstract: | |
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| Keywords: | Keywords: Brucella 26 kDa outer membrane protein tagged vaccine strain |
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