【作者】 张旭志； 吕志林； 李秋芬； 马绍赛； 崔毅；

【Author】 Xuzhi Zhang;Zhilin Lv;Qiufen Li;Shaosai Ma;Yi Cui;Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences;

【机构】 中国水产科学研究院黄海水产研究所；

【摘要】 <正>结合环介导等温扩增(LAMP)和电化学伏安检测技术,实验建立一种目标基因甄别新方法。成功的LAMP反应将把游离态的三磷酸2-脱氧鸟苷(d GTP,反应物之一)组装到链状产物DNA中去,结果引起该物质在碳纳米管阵列电极上氧化电流的显著下降。因此,这种伏安响应变化可以用来表征LAMP结果,进而指示出模板DNA中目标基因的信息。大肠埃希氏菌普通菌株中的malB基更多还原

【Abstract】 We described here a novel strategy for discriminating target gene by integrating loop-mediated isothermal amplification(LAMP) and differential pulse voltammetry(DPV). After a successful LAMP reaction, the anodic peak current(ipa) of the free 2’-deoxyguanosine 5’-triphosphate(d GTP) decreased remarkably at a carbon nanotubes array working electrode, owing to the consumption of free d GTP as one of reactive substrates. Thus, the change of current response was used to characterize the result of LAMP reaction. And hence the presence of the target gene in template DNA could be discriminated easily. The mal B gene extracted from Escherichia coli cells was tested as a model. After the reaction for 30 min, the LAMP mix was scanned directly. Then the information of the target gene in 0.8 picogram template DNA was obtained accurately. The result was in good accordance with that obtained with optical-based methods(gel electrophoresis and fluorescent dye). The new strategy has the advantages of being very simple to perform, rapid response, elimination of post-amplification processing, avoidance of auxiliary reagents and low cost(there was almost no cost for the detection step). Therefore, it was quite promising for use in miniaturized devices and in the development of point-of-care applications.更多还原