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| 10种常见细菌基因检测膜条的研制 | |
| 引用本文： | 朱飞舟,陈利玉,刘新发,田奇,谈成,欧阳方丹,吴正理,陈汉春.10种常见细菌基因检测膜条的研制[J].微生物学杂志,2011,31(4):58-63. |
| 作者姓名： | 朱飞舟 陈利玉 刘新发 田奇 谈成 欧阳方丹 吴正理 陈汉春 |
| 作者单位： | 1. 中南大学生物科学与技术学院生物化学系,湖南长沙,410013 2. 中南大学基础医学院医学微生物学系,湖南长沙,410013 3. 中南大学生物科学与技术学院生物实验中心,期南长沙,410013 |
| 基金项目： | 湖南省科技计划(2009SK3192); 教育部新教师基金(20090162120028); 中南大学贵重仪器设备开放共享基金(2011-29) |
| 摘 要： | 以16S rRNA基因为检测靶基因,设计10种常见细菌的DNA探针,将探针固定于硝酸纤维素膜条;PCR扩增细菌的16S rRNA基因片段并标记生物素后与膜条杂交;采用碱性磷酸酶标记的链亲和素检测生物素标记,以NBT/BCIP显色。该膜条不仅能单独检测10种细菌中的任何一种,也能同时检测5种细菌。该方法具备高通量、低成本、快速、准确等特点,具有良好的临床应用前景。 |
| 关 键 词： | 基因芯片 细菌 16S rRNA |
Development of Membrane Strip for Gene Detection of 10 Common Bacteria |
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| ZHU Fei-zhou,CHEN Li-yu,LIU Xin-f,TIAN Qi,TAN Cheng,OUYANG Fang-dan,WU Zheng-li,CHEN Han-chun.Development of Membrane Strip for Gene Detection of 10 Common Bacteria[J].Journal of Microbiology,2011,31(4):58-63. | |
| Authors: | ZHU Fei-zhou CHEN Li-yu LIU Xin-f TIAN Qi TAN Cheng OUYANG Fang-dan WU Zheng-li CHEN Han-chun |
| Affiliation: | ZHU Fei-zhou1,CHEN Li-yu2,LIU Xin-fa3,TIAN Qi1,TAN Cheng1,OUYANG Fang-dan1,WU Zheng-li1,CHEN Han-chun1(1.Dept.of Biochem.,School of Biol.Sci.& Technol.,2.Dept.of Pathog.Microbiol.,School of Basic Med.,3.Biol.Exper.Ctr.,Sch.of Biol.Sci.& Technol.,Zhongnan Uni.Changsha,410013) |
| Abstract: | 16S rRNA gene was taken as test target to design DNA probes for detecting 10 common bacteria then fixed the probes on nitrocellulose membrane strip.16S rRNA gene fragments of the 10 kinds of bacteria were amplified by PCR and labeled with biotin,then hybridized with the membrane strip.The biotin label was checked by SA-AP binding and developed with NBT/BCIP.This membrane strip could detect independently not only anyone of the 10 kinds of bacteria,but also 5 kinds of bacteria at the same time.This method pos... |
| Keywords: | gene chip bacteria 16S rRNA |
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