A Sweet Spot for Molecular Diagnostics: Coupling Isothermal Amplification and Strand Exchange Circuits to Glucometers.
A Sweet Spot for Molecular Diagnostics: Coupling Isothermal Amplification and Strand Exchange Circuits to Glucometers.
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DOI：
10.1038/srep11039
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发表时间：
2015-06-08
影响因子：
4.6
通讯作者：
Li B
中科院分区：
文献类型：
--
作者：
Du Y;Hughes RA;Bhadra S;Jiang YS;Ellington AD;Li B
链交换核酸电路可用于将等温核酸扩增产物转化为可在现成的血糖仪上可读的信号。环介导等温扩增（LAMP）受到非特异性产物积累的限制，但核酸电路可用于探测和区分特异性扩增子。通过将这种高温等温扩增方法与热稳定转化酶相结合，我们可以直接将中东呼吸综合征冠状病毒和扎伊尔埃博拉病毒模板转化为葡萄糖信号，灵敏度低至20-100拷贝/μl，相当于阿摩尔（或低zepto-mole）。来自细胞裂解物和合成模板的病毒甚至可以在痰或唾液中容易地扩增和检测。在病毒扩增子上协调触发的或门进一步保证了故障安全病毒检测。该方法描述了加速即时护理应用的潜力，因为生物样品可以应用于换能器，然后换能器将直接与现成的批准的医疗设备连接。
Strand exchange nucleic acid circuitry can be used to transduce isothermal nucleic acid amplification products into signals that can be readable on an off-the-shelf glucometer. Loop-mediated isothermal amplification (LAMP) is limited by the accumulation of non-specific products, but nucleic acid circuitry can be used to probe and distinguish specific amplicons. By combining this high temperature isothermal amplification method with a thermostable invertase, we can directly transduce Middle-East respiratory syndrome coronavirus and Zaire Ebolavirus templates into glucose signals, with a sensitivity as low as 20–100 copies/μl, equating to atto-molar (or low zepto-mole). Virus from cell lysates and synthetic templates could be readily amplified and detected even in sputum or saliva. An OR gate that coordinately triggered on viral amplicons further guaranteed fail-safe virus detection. The method describes has potential for accelerating point-of-care applications, in that biological samples could be applied to a transducer that would then directly interface with an off-the-shelf, approved medical device.
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