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个人电脑和手机向基因诊断系统的转变。

Transformation of personal computers and mobile phones into genetic diagnostic systems.

作者信息

Walker Faye M, Ahmad Kareem M, Eisenstein Michael, Soh H Tom

机构信息

Department of Chemistry and Biochemistry, University of California , Santa Barbara, California 93106, United States.

出版信息

Anal Chem. 2014 Sep 16;86(18):9236-41. doi: 10.1021/ac5022419. Epub 2014 Aug 31.

Abstract

Molecular diagnostics based on the polymerase chain reaction (PCR) offer rapid and sensitive means for detecting infectious disease, but prohibitive costs have impeded their use in resource-limited settings where such diseases are endemic. In this work, we report an innovative method for transforming a desktop computer and a mobile camera phone--devices that have become readily accessible in developing countries--into a highly sensitive DNA detection system. This transformation was achieved by converting a desktop computer into a de facto thermal cycler with software that controls the temperature of the central processing unit (CPU), allowing for highly efficient PCR. Next, we reconfigured the mobile phone into a fluorescence imager by adding a low-cost filter, which enabled us to quantitatively measure the resulting PCR amplicons. Our system is highly sensitive, achieving quantitative detection of as little as 9.6 attograms of target DNA, and we show that its performance is comparable to advanced laboratory instruments at approximately 1/500th of the cost. Finally, in order to demonstrate clinical utility, we have used our platform for the successful detection of genomic DNA from the parasite that causes Chagas disease, Trypanosoma cruzi, directly in whole, unprocessed human blood at concentrations 4-fold below the clinical titer of the parasite.

摘要

基于聚合酶链反应(PCR)的分子诊断技术为检测传染病提供了快速且灵敏的方法,但高昂的成本阻碍了其在这类疾病流行的资源有限地区的应用。在这项研究中,我们报告了一种创新方法,可将台式计算机和移动摄像手机(这两种设备在发展中国家已很容易获得)转变为高灵敏度的DNA检测系统。通过使用控制中央处理器(CPU)温度的软件将台式计算机转变为实际的热循环仪,实现了高效PCR,从而完成了这一转变。接下来,我们通过添加一个低成本滤光片将手机重新配置为荧光成像仪,使我们能够对PCR扩增产物进行定量测量。我们的系统灵敏度极高,能够定量检测低至9.6阿托克的目标DNA,并且我们证明其性能与先进的实验室仪器相当,而成本仅为后者的约1/500。最后,为了证明其临床实用性,我们利用我们的平台成功地直接从全血、未经处理的人体血液中检测出了导致恰加斯病的寄生虫——克氏锥虫的基因组DNA,其浓度比该寄生虫的临床滴度低4倍。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f0f3/4165218/bf49a3d66a51/ac-2014-022419_0006.jpg

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