Microbead-based rolling circle amplification in a microchip for sensitive DNA detection
Microbead-based rolling circle amplification in a microchip for sensitive DNA detection
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DOI：
10.1039/b927460j
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Kitamori, Takehiko
中科院分区：
文献类型：
--
作者：
Sato, Kae;Tachihara, Atsuki;Kitamori, Takehiko
在食品工业、环境和临床环境中，DNA靶标的灵敏检测和定量是确保无污染食品、监测环境和防治疾病的最重要问题。选择性探针与强大的扩增技术相结合，因此是主要的兴趣。在这项研究中，我们着手创建一个集成的微量化学芯片，受益于微流控芯片技术的灵敏度和强大的检测方法，由挂锁探针和滚环扩增（RCA）联合提供。在这里，我们将挂锁探针和RCA集成到微芯片中。该芯片在微通道中使用固相捕获，以实现洗涤循环并减少分析面积，并采用珠上RCA进行单分子扩增和检测。考察了试剂浓度和锁式探针用量对检测结果的影响，论证了锁式探针检测沙门氏菌的可行性。
The sensitive detection and quantification of DNA targets in the food industry and in environmental and clinical settings are issues of utmost importance in ensuring contamination-free food, monitoring the environment, and battling disease. Selective probes coupled with powerful amplification techniques are therefore of major interest. In this study, we set out to create an integrated microchemical chip that benefits from microfluidic chip technology in terms of sensitivity and a strong detection methodology provided jointly by padlock probes and rolling circle amplification (RCA). Here, we have integrated padlock probes and RCA into a microchip. The chip uses solid phase capture in a microchannel to enable washing cycles and decrease analytical area, and employs on-bead RCA for single-molecule amplification and detection. We investigated the effects of reagent concentration and amount of padlock probes, and demonstrated the feasibility of detecting Salmonella.