| 传统DNA探针实验方法的简化及应用 |
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| 引用本文： | 李银太,陈民钧.传统DNA探针实验方法的简化及应用[J].军事医学科学院院刊,1989,13(2):121-124. |
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| 作者姓名： | 李银太 陈民钧 |
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| 作者单位： | 军事医学科学院微生物流行病研究所，军事医学科学院微生物流行病研究所，军事医学科学院微生物流行病研究所，军事医学科学院微生物流行病研究所，中国医学科学院北京协和医院 |
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| 基金项目： | 国家自然科学基金，军事医学科学院基金 |
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| 摘 要： | 本文报告一种适于普通实验室开展DNA探针工作的实验方法.我们用酚-氯仿抽提法制备探针供体菌重组质粒DNA,用透析袋电泳洗脱法从凝胶中回收酶切目的片段,用缺口翻译技术制备α-~(32)PdNTP标记探针,用低脂奶粉和6SSC(1×SSC:0.15M NaCl,0.015M柠檬酸三钠,pH7.0)代替Denhardt试剂等获得了满意的菌落原位杂交和Southern blot杂交结果.此外,我们将传统实验方法规定的时间相应缩短,从而有可能在15～24h内对痢疾腹泻患者标本作出明确诊断.这些方法可供基层实验室应用DNA探针开展传染病诊断和流行病学调查.
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| 关 键 词： | DNA探针 菌落原位杂交 痢疾杆菌 |
SIMPLIFICATION OF CONVENTIONAL DNA PROBE TECHNOLOGY AND ITS APPLICATION |
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| Li Yintai,Guo Zhaobiao,Lin Wanming,Wang Jin,Chen Minjun Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences.SIMPLIFICATION OF CONVENTIONAL DNA PROBE TECHNOLOGY AND ITS APPLICATION[J].Bulletin of the Academy of Military Medical Sciences,1989,13(2):121-124. |
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| Authors: | Li Yintai Guo Zhaobiao Lin Wanming Wang Jin Chen Minjun Institute of Microbiology and Epidemiology Academy of Military Medical Sciences |
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| Abstract: | A method suitable for the detection of DNA probe in common laboratories is described.Recombinant plasmid of the donor bacteria for DNA probe was prepared by phenol-chloro-form extraction.The DNA fragment was recovered by means of electrophoretic elution with dialysis bag.~(32)PdNTP labelled probe was prepared by nick translation technique.In the DNA probe experiments,we had used low-fat milk powder as a substitute for Denhardt's reagent and obtained satisfactory results of in situ hybridization.In addition,diagnostic results of stool specimens of dysentery patients could be obtained in 15-24 hrs.This method may be used in the clinical diagnosis of various infectious diseases and epidemiological survey in grass root units laboratories. |
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| Keywords: | DNA probe colony in situ hybridization Shigella |
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