痰标本；结核分枝杆菌DNA；抽提;显色法；耐药基因；基因芯片

不同样本处理方法对显色法芯片检测结核分枝杆菌利福平和异烟肼耐药基因的影响

http://www.100md.com 《实用医技杂志》 2006年第21期

痰标本；结核分枝杆菌DNA；抽提;显色法；耐药基因；基因芯片,,],痰标本；结核分枝杆菌DNA；抽提;显色法；耐药基因；基因芯片,1材料和方法,2结果,3讨论,参考文献：

     [摘要] 目的:选择一种较为理想的从痰标本中直接抽提结核分枝杆菌DNA的方法，用于显色法芯片检测结核分枝杆菌利福平(RFP)和异烟肼(INH)的耐药基因。方法:选择3份含结核分枝杆菌分别为4.325×103、6.857×104、5.356×105 Copies/ml的痰液，分别用Chelex 100树脂法、煮沸法、蛋白酶K消化异丙醇沉淀法、蛋白酶K消化酚/氯仿抽提法、碱裂解法、异硫氰酸胍裂解法、盐酸胍裂解法、离心柱法抽提结核分枝杆菌DNA作多重聚合酶链反应，显色法芯片检测结核分枝杆菌RFP和INH耐药基因。结果:离心柱法检测结果阳性，其余方法检测结果均为阴性，离心柱法检测灵敏度为2.743×103 Copies/ml。结论:离心柱法是一种较为理想的从痰标本中直接抽提结核分枝杆菌DNA的方法,可用于显色法芯片检测结核分枝杆菌RFP和INH耐药基因。

    [关键词] 痰标本；结核分枝杆菌DNA；抽提;显色法；耐药基因；基因芯片

    Effect of Varied Sample Treatment Procedures on Detection of the Resistance Genes of Rifampin and Isoniazid in Mycobacterium Tuberculosis DNA by Using Coloration Genes Chips

    WANG Wanxiang,GUO Naizhou，YAN Aihua,MA Li,ZHOU Buquan,MA Da,JING Fengxiang,TANG Xiangrong,ZHUANG Yan

    (The First People's Hospital of Yancheng City,Yancheng，Jiangsu 224006,China；The Second People's Hospital of Yancheng，Yancheng,Jiangsu 224002,China;Shanghai Micro System and Information Technique Graduate School of China Academy of Sciences,Shanghai 200050,China;Jingjiang College of Jiangsu University,Zhenjiang,Jiangsu 212013,China)

    Abstract:Objective To choose a more ideal method to directly extract My cobacterium tuberculosis DNA from sputum sample to detect the resistance genes of rifampin (RFP) and isoniazid (INH) in Mycobacterium tuberculosis by using Coloration gene chips.Methods Choose three sputum samples whose Mycobacterium tuberculosis respectively were 4.325×103,6.857×104 and 5.356×105 Copies/ml.Respectively use chelex 100 resinate,boiling extraction ......

百拇医药网 http://www.100md.com/html/DirDu/2007/03/02/37/79/32.htm

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