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DNA固定技术对结核分枝杆菌PCR产物杂交结果影响的研究

DNA fixation technique on influence of hybridization results of PCR products of M.tuberculosis

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【作者】 李传友; 马屿; 邢爱英; 古淑香; 贾红艳;

【Author】 Li Chuanyou,Ma Yu,Xing Aiying,et al. Beijing TB & Thoracic Tumor Institute,Beijing 101149

【机构】 北京市结核病胸部肿瘤研究所!101149;

【摘要】 目的 比较各种DNA 固定方法对结核分枝杆菌PCR 产物杂交结果的影响。方法 选取结核分枝杆菌PCR 扩增产物并点于尼龙膜上,用紫外线、加热和碱液三种方法固定DNA,然后以PCR 产物作探针用增强化学发光标记试剂进行标记,并对三种不同方法固定的PCR 产物杂交分析。结果 结核分枝杆菌DNA固定方法对PCR 扩增产物的检测结果具有较大的影响,碱液固定具有较好的敏感性和重复性。结论 各地结核分枝杆菌PCR 实验室可考虑用碱液法对PCR 产物固定进而进行杂交分析

【Abstract】 Objective To compare influence of various DNA fixation methods on hybridization of PCR products of M.tuberculosis.Methods PCR products of M.tuberculosis were obtained,then diluted into different concentration and added to the nylon membrane.PCR products on the nylon membrane were fixed with UV crosslinking,oven baking and alkali fixation method,then hybridized and analysed.Results It was found that DNA fixation method had a great influence on detection of PCR products of M.tuberculosis.Alkali fixation had a good sensitivity and reproducibility.Conclusion All the results suggested that PCR laboratory of M.tuberculosis could consider using alkali fixation to PCR products,then hybridized and detected.

【关键词】 DNA固定; 分枝杆菌; 结核; PCR; 杂交;
【Key words】 DNA fixation M.tuberculosis PCR Hybridization;
【基金】 北京市自然科学基金
  • 【文献出处】 中国防痨杂志 ,BUCCETIN OF CHINESE ANTITUBERCULOSIS ASSOCLATION , 编辑部邮箱 ,1999年04期
  • 【分类号】R378.911
  • 【下载频次】52
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